Spectroscopic Techniques and

their Applications in Chemistry

 Introduction
Spectroscopy is a branch of science that involves study on interaction of
electromagnetic radiation with matter.

• One of the nature’s most beautiful sights is rainbow. But, it was Isaac
newton who first split sunlight using a prism to create the color of
rainbow.

Newton’s prism experiment

 Electromagnetic Radiation
• Electromagnetic radiation is a simple harmonic wave of electric and
magnetic fields oscillating orthogonal to each other

• Visible light is electromagnetic radiation

 Energy, Wavelength and Frequency

ℎ𝑐 E = Energy
𝐸 = h = Planck's constant, 6.626 x10-34 m2 kg s-1
λ ν = Frequency
𝑐 c = Velocity of light, 3.0 x108 m/s
𝐸 = ℎν; ν = λ = Wavelength
λ

E  ; E  −;   −
Different Regions of EM Spectra
 Types of Spectroscopy

Absorption spectroscopy Emission spectroscopy

• Transition takes place • Transition takes place

from lower to higher from higher to lower
energy levels energy levels

• molecules have discrete (quantized) energy levels

• electromagnetic radiation is absorbed when the energy
of photon corresponds to the difference in energy
between two states.
In order to interact with the electromagnetic radiation, the molecules must
have some electric or magnetic effect that could be influenced by the
electric or magnetic components of the radiation.

(A) Rotation of a water molecule around its center of mass.

(B) The change in the dipole moment as a result of rotation.
 Broadening of Peaks

• Intrinsic broadening of peaks arises from the Heisenberg’s uncertainty

principle, which indicates that shorter the lifetime of a state, the more
uncertain is its energy. Molecular transitions have finite lifetimes, therefore
their energy is not exact. If Δt is the lifetime of a molecule in an excited state,
the uncertainty in the energy of the states is given by:

Δ𝐸 × Δ𝑡 ≥ ℎ/4𝜋

• Molecules in a liquid or gaseous sample are in motion and keep colliding with
each other, which influence the vibrational and rotational motions of the
molecules thereby causing broadening. Two atoms/molecules coming in
close proximity will perturb the electronic energies, at least those of the
outermost electrons resulting in broadening of electronic spectra.
 Utility of Spectroscopy in Chemistry

UV-Vis: electronic transitions

- gives information about p-bonds and conjugated systems

Infrared: molecular vibrations (stretching, bending)

- identify functional groups

Radiowaves: nuclear spin in a magnetic field (NMR)

- gives a map of the H and C framework
Electronic Spectroscopy
We see the complementary color of the wavelength is absorbed
by a material.

Color of Color
absorbed light  observed

violet 400 nm yellow

blue 450 orange
blue-green 500 red
yellow-green 530 red-violet
yellow 550 violet
orange 600 blue-green
red 700 green
 Electronic Transitions

UV-vis light causes electrons in lower energy molecular orbitals to be

promoted to higher energy molecular orbitals.

Wavelength range for UV light: 200-400 nm

Wavelength range for visible light: 400-800 nm
Different Types of Electronic Transitions
 Examples of Different Electronic Transitions

In all compounds other than alkanes, the electrons may undergo several possible
transitions of different energies. Some of the most important transitions are
Single Beam
Spectrophotometer

Double Beam
Spectrophotometer
UV Absorption Spectra of Phenol in Different
Solvents

• λmax is the
wavelength at
which absorbance
is maximum.

• Note that the

spectrum becomes
broad when the
solvent is more
interacting in nature.
 Few Terminologies

• Bathochromic shift (red shift)—a shift to lower energy or longer

wavelength
• Hypsochromic shift (blue shift)—a shift to higher energy or shorter
wavelength
• Hyperchromic effect—an increase in intensity
• Hypochromic effect—a decrease in intensity
 Lambert-Beer Law
Absorption of the light depends on the number of molecules and thickness
of the sample cell.

There is a linear relationship between absorbance and concentration.

A=cl

when l = 1 cm

percentage transmittance:
 A = absorbance/ optical density = log(Io/I)

Io is initial intensity of radiation and I is intensity after it passes

through an absorbing solution

 = molar absorptivity or extinction coefficient

 is a proportionality constant for a specific absorbance
of a substance at a given wavelength

c = concentration (M, mol/L)

l = sample path length (cm)

• Note that Beer-Lambert law is applicable for dilute solutions. At higher

concentrations, the molecules come in close proximity thereby influencing
their electronic properties. ε can practically be assumed to be a constant at
low concentrations only for a compound at a particular wavelength. ε
depends on the concentration of the compound when the concentration is
high.
 Effect of Conjugation on Absorption Wavelength

(a)

(b)

benzene naphthalene anthracene tetracene

 Chromophores
Chromophores are the light absorbing part of a molecule

Some natural dyes

indigo beta carotene

lycopene

chlorophyll
 Woodward-Fieser Rules for Calculating λmax of conjugated
Molecules
conjugated diene: base value 217 nm

homonuclear diene heteronuclear diene

(cisoid) (transoid)
 Examples
(a) (b)

(c) (d)
 Applications of Electronic Spectroscopy

• Quantitative analysis: Determination of unknown concentrations by Beer-

Lamberts law.

• Detection of impurities: It is one of the best method for detecting impurities

in organic solvents. For example, benzene is the most common impurity in
cyclohexane. Benzene can be detected by its absorption band at 255 nm.

• Study kinetics of chemical reaction: Fix the wavelength of ether reactant or

product and measure absorbance at different time intervals. Plot
absorbance vs time. Slope contains information about rate constant.