Modern Chemical

Instrumentation

UNIT-1
Spectroscopy
 Basics
• Spectroscopy deals with the study of interaction of
electromagnetic radiation with matter.
• Electromagnetic radiation is a simple harmonic wave of
electric and magnetic fields fluctuating orthogonal to each
other

An electromagnetic wave showing orthogonal electric and magnetic

components (A); a sine wave (B)
 Basics
• A simple harmonic function can be represented by a sine
wave; 𝑦 = 𝐴 sin𝜃

• This equation can therefore be written in terms of angular

velocity, ω.
• 𝑦 = 𝐴 sin(𝜔𝑡) » 𝑦=𝐴 sin(2𝜋ν𝑡) » 𝑦=𝐴 sin(2𝜋ν𝑧/𝑐)
where, z = displacement in time t and c is the velocity of the
electromagnetic wave

• If the wave completes ν cycles/s and the wave is travelling

with a velocity c metres/ sec, then the wavelength of the wave
must be 𝑐 / ν metres.
• 𝑦 = 𝐴 sin(2𝜋𝑧/𝜆)
 Basics
• Energy of an EM radiation:
𝐸 = ℎν = ℎ𝑐 / 𝜆
where h is Planck’s constant
(6.626 × 10-34 m2·kg·s-1)

● Based on the energy, electromagnetic radiation has been

divided into different regions.
● Not convenient to write such small values of energy; the
therefore written in electronvolts (eV). One electronvolt
equals 1.602 × 10-19 Joules. The energy range of the visible
spectrum is ~1.6 – 3.1 eV.
 Electromagnetic spectrum
● The region of electromagnetic spectrum human beings can
see is called visible region or visible spectrum.
● The visible region constitutes a very small portion of the
electromagnetic spectrum and corresponds to the wavelengths of
~400 – 780 nm. The energy of the visible spectrum therefore ranges
from ~2.5 × 10-19 to ~5 × 10-19 Joules.

•
Visible spectrum
 Quantization of Energy
● According to Max Planck, atoms and molecules can absorb or
emit the energy in discrete packets, called quanta/ quantum.
● The quantum for electromagnetic energy is called a photon
which has the energy given by equation; 𝐸 = ℎν = ℎ𝑐/ 𝜆
● A molecule can possess energies in different forms such as
vibrational energy, rotational energy, electronic energy, etc.
● The electrons reside in different orbits/orbitals surrounding
the nucleus, in discrete electronic energy levels of atoms.
● In
• much the same way, rotational and vibrational energy
levels of molecules are also discrete.
● A molecule can jump from one energy level to another by
absorbing or emitting a photon of energy that separate the two
energy levels
Quantization of Energy
• Transitions of a molecule between energy levels, E1 and E2
by absorbing/emitting the electromagnetic radiation .
 EM spectrum & the atomic/molecular processes:
• Molecules undergo processes like rotation, vibration,
electronic transitions, and nuclear transitions.
• The energies underlying these processes correspond to
different regions in the electromagnetic spectrum
• Radiofrequency waves: Radiofrequency region has very
low energies that correspond to the energy differences in
the nuclear and electron spin states. These frequencies,
therefore, find applications in nuclear magnetic resonance
and electron paramagnetic resonance spectroscopy.
• Microwaves: Microwaves have energies between those of
radiofrequency waves and infrared waves and find
applications in rotational spectroscopy and electron
paramagnetic resonance spectroscopy.
EM spectrum & the atomic/molecular processes:
 EM spectrum & the atomic/molecular processes:
• Infrared radiation: The energies associated with molecular
vibrations fall in the infrared region of electromagnetic
spectrum. Infrared spectroscopy is therefore also known as
vibrational spectroscopy and is a very useful technique for
functional group identification in organic compounds.
• UV/Visible region: UV and visible regions are involved in
the electronic transitions in the molecules. The
spectroscopic methods using UV or visible light therefore
come under ‘Electronic spectroscopy’.
• X-ray radiation: X-rays are high energy electromagnetic
radiation and causes transitions in the internal electrons of
the molecules.
Interaction of EM radiation with matter:
• 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.

• In NMR spectroscopy, for example, the nuclear spins have

magnetic dipoles aligned with or against a huge magnetic field.
Interaction with radiofreqency of appropriate energy results in the
change in these dipoles.
• Rotations of a molecule having a net electric dipole moment, such
as water will cause changes in the directions of the dipole and
therefore in the electrical properties. There are changes in the y-
component of the dipole moment due to rotation of water
molecule.
• Fig A shows rotation of a water molecule around its centre of mass
• Fig B shows the change in the dipole moment as a result of rotation
Interaction of EM radiation with matter:
Interaction of EM radiation with matter:
• Vibrations of molecules can result in changes in electric dipoles
that could interact with the electrical component of the
electromagnetic radiation.
• Figure C shows the change in dipole moment of water due to
asymmetric stretching vibrations of O—H bond
• Electronic transitions take place from one orbital to another.
Owing to the differences in the geometry, size, and the spatial
organization of the different orbitals, an electronic transition
causes change in the dipole moment of the molecule.
• Figure D shows an electronic transition from π to π* orbital and
the geometry of the two orbitals.

• The above examples suggest that a change in either electric or

magnetic dipole moment in a molecule is required for the
absorption or emission of the electromagnetic radiation.
 Absorption peaks and line widths
• Absorption of radiation is the first step
• Absorption spectra are observed for the electronic,
rotational, and vibrational spectroscopy.
• A transition between states takes place if the energy
provided by the electromagnetic radiation equals the energy
gap between the two states.
• Δ𝐸 = ℎ𝜐 = ℎ𝑐/ 𝜆
• This implies that the molecule precisely absorbs the
radiation of wavelength, λ and ideally a sharp absorption line
should appear at this wavelength
• However, in practice, the absorption lines are not sharp but
appear as fairly broad peaks.
Absorption lines

• An idealized
spectrum for a
single wavelength
transition (A) and
an experimentally
obtained spectrum
(B)
 Reasons for peak broadening
• Instrumental factors: The slits that allow the incident light to
impinge on the sample and the emerging light to the detector
have finite widths. Consider that the transition occurs at
wavelength, λt. When the wavelength is changed to λt + Δλ or
λt – Δλ, the finite slit width allows the radiation of wavelength,
λt to pass through the slits and a finite absorbance is observed at
these wavelengths.

• Sample factors: Molecules in a liquid or gaseous sample are in

motion and keep colliding with each other. Collisions 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. Motion of molecules undergoing transition also causes
shift in absorption frequencies, known as Doppler broadening.
 Reasons for peak broadening
• Intrinsic broadening: Intrinsic or natural broadening arises
from the Heisenberg’s uncertainty principle which states that
the 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

• Noise is the manifestation of the random weak signals

generated by the instrument electronics. To identify the
sample peaks clear of the noise, the intensity of the sample
peaks has to be at least 3-4 times higher than the noise. A
better signal-to-noise ratio should be obtained and the noise
should be cancelled out.
Lambert Beer Law
This helps us to define the relationship of the intensity of visible UV
radiation with the exact quantity of substance present.
The Beer-Lambert law states that for a given material sample
path length and concentration of the sample are directly
proportional to the absorbance of the light.
A = εlc
Lambert Beer Law
Lambert Beer Law
Lambert Beer Law
Lambert Beer Law
Lambert Beer Law: Limitations
o The linearity of the Beer-Lambert law is limited by chemical
and instrumental factors.
o The electromagnetic radiation should be monochromatic.
o The light beam should not be scattered.
o The solution should be diluted.

Causes of non-linearity occurs in the following conditions:

• Deviations in absorptivity coefficients at high concentrations (>
0.01M) due to electrostatic interactions between molecules in close
proximity
• Scattering of light due to particulates in the sample
• Fluoresecence or phosphorescence of the sample
• Changes in refractive index at high analyte concentration
• Shifts in chemical equilibria as a function of concentration
• Non-monochromatic radiation
• Stray light
Instrumentation
Instrumentation: Radiation Sources
Instrumentation: Radiation Sources

Emission spectrum of a
commercial deuterium arc lamp
Instrumentation: Monochromators
Instrumentation: Monochromators
Instrumentation: Monochromators
Instrumentation: Monochromators
Cross-section diagram of a diffraction
grating showing diffraction of a single
beam of light. Symbols: i : angle of
incidence, u : angle of diffraction (or
reflectance), b : blaze angle of
the grating, d : grating spacing.
Prism used as an
order sorter for a
grating
monochromator.
Instrumentation: Detectors