Electromagnetic spectrum: The entire range of electromagnetic spectrum is given by cosmic

rays, gamma rays, X-rays, ultra-violet radiation, visible light, infrared radiation, microwaves and
radio waves in the increasing order of wavelength and decreasing order of frequency

Type of Radiation Frequency (Hz) Wavelength (λ)

Range Range
Gamma-rays 1020 – 1024 < 10-12 m
X-rays 1017 – 1020 1 nm – 1 pm
Ultraviolet 1015 – 1017 400 nm – 1 nm

Visible 4 x 1014 – 7.5 x 1014 750 nm – 400 nm

Near-infrared 1 x 1014 – 4 x1014 2.5 μm – 750 nm

Infrared 1013 – 1014 25 μm – 2.5 μm

Microwaves 3 x 1011 – 1013 1 mm – 25 μm

Radio waves < 3 x 1011 > 1 mm

Absorption of radiation: Beer-Lambert’s law
Whenever radiation interacts with matter, three things happen: (i) a part of it is absorbed (ii) a part
of is transmitted and (iii) the remaining part is reflected. For transparent solutions, the
reflected/scattered part is almost negligible. Most of it is either absorbed or transmitted.
Beer-Lambert law is stated as ‘when a monochromatic light is passed through a transparent and
coloured solution, the intensity of light absorbed is directly proportional to the concentration of the
absorbing species (C) and the length of the solution (X).
i.e. A  C.X
In general Beer-Lambert law is usually written as:

Here A is the measured absorbance,

ε is molar extinction coefficient which depends on the wavelength,
X is the path length of light (or length of solution) and
C is the molar concentration of the analyte.
Based on this Beer-Lambert’s law, if the absorbance of a sample solution at a particular wave length
is known, then the concentration of it can be estimated.
UV-VIS SPECTROSCOPY

UV-Visible spectroscopy is a technique that measures the absorption or

transmission of light by a sample in the ultraviolet (190-400 nm) and visible
(400-800 nm) regions of the electromagnetic spectrum.

It is based on the principle that molecules undergo electronic transitions

when they absorb light of specific wave lengths.

The amount of light absorbed or transmitted depends on the sample

composition and concentration, as well as the path length of light travelled
through the sample
Electronic transitions:
 An electronic transition is the movement of an electron from one
energy level to another within a molecule, usually in response to the
absorption or emission of light.
 The energy change is associated with an electronic transition and it
provides information on the structures and properties of molecule,
such as its colour and absorption spectrum.
The diagram shows the relative energies required for various types of electronic transitions
(i)   * Transition:
 This occurs when an electron in a sigma () bonding orbital is
excited to a sigma star (*) anti bonding orbital.

 This requires a high amount of energy and usually occurs in the

vacuum ultraviolet region (below 200 nm)

 This type of transition is observed in molecules with only sigma

bonds, such as H2, CH4 and NH3.
ii) n  * Transition:
 This occurs when an electron in antibonding (n) orbital, such as a
lone pair is excited to a sigma star (*) anti bonding orbital.

 This requires less energy than a   * transition and usually

occurs in the far UV region (200-300 nm)

 This type of transition is observed in molecules with hetero

atoms that have lone pairs such as NH3, H2O and CH3Cl
iii)   * Transition:
 This occurs when an electron in pi () bonding orbital is excited to a pi star
(*) antibonding orbital.

 This requires even less energy than a n  * transition and usually occurs
in the near UV region (300-400)

 This type of transition is observed in molecules with double or triple bonds,

such as C2H4, C2H2 and CO2.
iv) n  * Transition:
 This occurs when an electron in a non-bonding (n) orbital is excited to a
pi star (*) anti bonding orbital.

 This requires least amount of energy among the four types of transitions
and usually occurs in the visible region (400 – 800 nm)

 This type of transition is observed in molecules with both heteroatoms

and double or triple bonds, such as H2O, NH2 and CH2O.
Instrumentation of UV-Visible spectroscopy

 A spectrophotometer is an instrument for measuring the

transmittance or absorbance of a sample as a function of
wavelength.

 It is also used in measurement of absorbance of a series of samples

at a single wavelength.

 UV Visible spectrophotometers are available in single or double

beam spectrophotometers.
(i) Single beam spectrophotometer: In this the light passes through the sample and
the detector in series and the absorbance is calculated by comparing the
intensity of the light before and after passing through the sample.

(ii)Double beam spectrophotometer: In this the light is split into two beams by a
beam splitter and one passes through the sample while the other passes through
a reference. The absorbance is calculated by comparing the ratio of the
intensities of two beams.

The essential components of a spectrophotometer (both single and double beam)

are shown schematically in the following figure
Block Diagram of UV Visible spectrophotometer
The important components of UV-Visible spectrophotometer

(a) Light source: It provides two lamps as sources of UV & Visible light.
(i) Deuterium lamp [D2] is used for UV region [200 to 400 nm]
(ii) Tungsten lamp is used for visible region [400 to 700nm]

Note: The generated heat from tungsten lamp may warm up the sample
and other components of the instrument, so that the lamp is often cooled with
an air blower.
(b) Filter:
Commonly used for reflection, transmission, absorption, and blocking UV
light or wavelengths, which they do without affecting, light within the visible
region.

(c) Monochromator:

 Monochromators are used to control the wavelength of light when needed.

 Monochromator resolves a broad band of spectrum into narrow band of

radiation with high spectral purity. prisms or diffraction gratings.
(d) Beam Spiltter:
 A split beam spectrophotometer contains a beam splitter which channels
light along a reference path and a sample path simultaneously to two
separate detectors

(e) Photo diode/Detectors:

 The transmitted light passing through the sample/reference is detected by a
photo diode.
 sA photodiode array in UV-Visible spectrophotometer, acquires data at
each wavelength by electrical scanning.
 photomultiplier tube (PMT) is one of the more common detectors used in
UV‑Visible spectroscopy.
(f) Amplifier and Recorder: The current coming from the detector is
amplified with the help of amplifier which is recorded.
Based on this data of transmitted light intensity, the absorbed light
intensity is obtained and a plot of wave length versus absorbance is
obtained as shown.
 INFRARED SPECTROSCOPY

Infrared Spectroscopy generally refers to the analysis of the interaction of a

molecule with infrared light

In IR spectroscopy, the samples can generally be analyzed in three ways: by

measuring reflection, transmission, and absorption.

The major use of infrared spectroscopy is to determine the functional

groups of molecules, relevant to both organic and inorganic chemistry

IR Spectroscopy detects frequencies of infrared light that are absorbed by a

molecule.

Molecules tend to absorb these specific frequencies of light since they

correspond to the frequency of vibration of bonds in the molecule
Regions of the Infrared spectrum:

Most of the bands that indicate what functional group is present are found in the
region from 4000 cm-1 to 1300 cm-1.

Their bands can be identified and used to determine the functional group of an
unknown compound.

Bands that are unique to each molecule, similar to a fingerprint, are found in the
fingerprint region, from 1300 cm-1 to 400 cm-1.

These bands are only used to compare the spectra of one compound to another
 Fundamental modes

Vibrations can involve either change in bond length (stretching) or bond

angle (bending).

Some bonds can stretch in-plane (symmetric stretching) or out-of-plane

(asymmetric stretching).

Bending vibrations can be either in-plane (as; scissoring, rocking) or out-of-

plane (as; wagging, twisting) bending vibrations.
1) Stretching vibrations for a diatomic molecule like H2O can be
represented as follows:

(a) Symmetric stretching: Terminal atoms move away from or towards each
other from the central atom, simultaneously.

(b) Aymmetric stretching: One atom moves away while the other atom moves
towards the central atom, simultaneously.
(ii) Bending vibrations for a methylene group (-CH2-) can be represented as
follows:

(a) Scissoring: Two adjacent atoms of a central atom move towards and away from
each other (like scissors) in the same plane.

(b) Rocking: Two adjacent atoms of a central atom move to the same side relative to
the central atom, in the same plane.

(c) Twisting: Out of plane bending of terminal atoms in opposite direction.

(d)Wagging: Out of plane bending of terminal atoms in same direction.

IR Spectroscopy Instrumentation
First a beam of IR light from the source is split into two and passed through the
reference and the sample.

Now both of these beams are reflected to pass through a splitter and then
through a detector.

Finally, the required reading is printed out after the processor deciphers the data
passed through the detector.
IR Spectroscopy Instrumentation
The main parts of the IR spectrometer are as follows:
1. Radiation source
2. Sample cells and sampling of substances
3. Monochromators
4. Detectors
5. Recorder
1. IR radiation sources
Various sources of IR radiations are: Nernst glower, Incandescent lamp,
Mercury arc, Tungsten lamp, Glober source, Nichrome wire

2.Sample cells and sampling of substances

IR spectroscopy has been used for the characterization of solid, liquid, or gas
samples.
(i) Solid – Various techniques are used for preparing solid samples such as
pressed pellet technique, solid run-in solution, solid films, mull technique, etc.

(ii) Liquid – Samples can be held using a liquid sample cell made of alkali
halides. Aqueous solvents cannot be used as they will dissolve alkali halides. Only
organic solvents like chloroform can be used.

(iii) Gas– Sampling of gas is similar to the sampling of liquids

3. Monochromators
Various types of monochromators are prism, gratings and filters.
Prisms are made of Potassium bromide, Sodium chloride or
Caesium iodide.
Filters are made up of Lithium Fluoride and Diffraction gratings
are made up of alkali halides.

4. Detectors
Detectors are used to measure the intensity of unabsorbed
infrared radiation.
Detectors like thermocouples, Bolometers, thermistors, Golay
cell, and pyro-electric detectors are used.
5. Recorders
Recorders are used to record the IR spectrum.