CH8491 – Instrumental Methods of Analysis 2019-2020

UNIT I
INTRODUCTION TO SPECTROSCOPICAL METHODS OF ANALYSIS

ANALYSIS

Analysis refers to the detailed examination of the elements and their structure.
Chemical analysis uses instruments and methods used to separate, identify, and quantify
matter. Analytical chemistry is the science of obtaining, processing, and communicating
information about the composition and structure of matter.

Analysis

Qualitative Quantitative
Analysis Analysis

Qualitative analysis
Qualitative analysis deals with the identification of elements or functional groups
present in a sample. In short, quantitative analysis answers, “What is it?”
Example: inorganic salt analysis (identification of cations and anions), organic elemental
analysis, organic functional group analysis.

Quantitative analysis
Quantitative analysis deals with the quantification of the substance. It tells you
how much substance is present. In short, quantitative analysis answers, “How much is
it?” Example: Volumetric analysis (titrations)

Methods of
Analysis

Classical Instrumental
Methods Methods

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Classical methods are the techniques, which forms the fundamentals of laboratory
practices. These are the traditional method of chemical analysis, which is still being used
by scientists. Classical method is cheaper and easily available for schools and industries.
It is sometimes called as "wet chemistry" where there are too many chemical reactions
are used to identify certain compounds. The classical method consumes more time than
the instrumental analysis. These methods are often labor intensive. The results are often
accurate and precise. Examples: Quantitative analysis performed by gravimetric and
volumetric (titrimetric) methods.

Instrumental Analysis
Instrumental methods are often called modern method of analysis. The reason is
that these analytical techniques use modern equipment’s such as computers and other
electronic equipment. Instrumental method is expensive because the machines are highly
specialized for a particular chemical analysis. Quantitative results are often more accurate
and precise than the classical methods. Precision is dependent less on the operator and
more on the instrument and sources of noise. Not all schools and colleges can afford
those machines. Machines used are quite sophisticated that experts are needed to operate
the machines to avoid system malfunctioning.
Examples: Qualitative analysis using Gas Chromatography, High Performance Liquid
Chromatography, NMR (Nuclear Magnetic Resonance) spectroscopy etc.

Advantages
 Less labor intensive
 Easy to automate
 Simultaneous multi component analysis
 Fast analysis
 Lower detection limits

Disadvantages
 Higher expense
 Harder to trouble shoot problems, more technical expertise

Spectroscopy
Spectroscopy is one of the most powerful tool used to derive information about
atomic and molecular structure. Spectroscopy is the branch of science dealing with the
study of interaction of electromagnetic radiation with matter. It involves the study of
radiation emitted, absorbed, transmitted or scattered by the matter under study. It
provides valuable information regarding chemical structure such as functional groups,

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unsaturation, bond strength, molecular structure such as molecular symmetry, bond

distances, bond angles etc.

Spectrometry
The term Spectrometry is used for the quantitative measurement of the intensity of one or
more wavelengths in the electromagnetic radiation. It is the method used for the study of
certain spectrums. Ion-mobility spectrometry, mass spectrometry are some examples of
spectrometry. In these cases, a spectrum does not necessarily mean a plot of intensity
versus frequency. For example, the spectrum for mass spectrometry is the plot between
intensity (number of incident particles) versus the mass of the particle. Spectrometers are
the instruments used in spectrometry. The operation of each type of instrument depends
on the form of spectrometry used in the instrument.

Difference between spectroscopy and spectrometry

S.no. Spectroscopy Spectrometry
1 Spectroscopy is the science of Spectrometry is the method used to
studying the interaction between acquire a quantitative measurement
matter and radiation of the spectrum (metry means
measurement).

2 Spectroscopy does not generate any Spectrometry is the practical

results. It is the theoretical approach of application where the results are
science generated

Electromagnetic radiation
Electromagnetic radiation (EMR) is a form of energy transmitted through space at
very high velocities. It requires no supporting media. It more readily passes in vacuum
than in a supporting medium like air. This energy has wave nature and is associated with
both electric and magnetic fields and hence, the name. Electric and magnetic field
oscillate perpendicular to each other and both are perpendicular to the direction of
propagation of wave.
The most obvious example of electromagnetic radiation is light which occupies
only a small region in a spectrum of electromagnetic radiation.

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Representation of electromagnetic radiation

Properties of electromagnetic waves

 Electromagnetic waves are composed of oscillating electric and magnetic fields at
right angles to each other and both are perpendicular to the direction of the
propagation of the wave.
 Do not require a medium for transmission.
 Travel at the speed of light (3 × 108 ms –1) in vacuum.
 Have no mass.
 Electromagnetic waves are transverse waves.
 Deflected by neither electric nor magnetic fields.
 Follow the laws of reflection and refraction
 Energy of electromagnetic waves is directly proportional to its frequency.
Electromagnetic Spectrum
The entire range over which the electromagnetic radiation exist is known as
electromagnetic spectrum. The electromagnetic spectrum covers an immense range of
wavelength. The various spectrum regions are

 Gamma rays
 X-Ray
 Visible
 Ultraviolet region
 Infrared region
 Microwave region
 Radio frequency region

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The electromagnetic spectrum (The visible portion is shown on an expanded scale to the right)

Major characteristics of various spectrum regions

Gamma ray region
This lies between 0.02 to 1 Å. The gamma rays are the shortest waves emitted by
atomic nuclei, involving energy changes of 109 to 1011 joules/gram atom.
X- Ray region
This lies between 1 to 10 Å. They are emitted or absorbed by movement of
electrons close to the nuclei of relatively heavy atoms, involve energy changes of the
order of ten thousand kilo joules.
Visible and ultraviolet region
The ultraviolet region ranges from 180 – 400 nm. The visible range extends from
400-750 nm.

Colors of visible light

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Infrared region
This region has been further divided in to the following sub-regions:
 Infrared (Near) – 0.7-2.5 µ
 Infrared – 2.5-15 µ
 Far infrared – 15-200 µ
All the three sub-region of infrared part of the electromagnetic spectrum are
associated with the change in molecular vibrations.
Microwave region
This region corresponds to the change in the rotation of molecules. It lies between
0.1mm to 1 cm wavelength. Separations between the rotational levels of molecules are of
the order of hundreds of joules per mole.
Radiofrequency region
The energy change involved in this region arises due to the reversal of a spin of
nucleus or electron. It lies between 10 m to 1 cm wavelength.

Dual Properties of Electromagnetic Radiation

Dual nature of light means light acts both as a particle and as a wave.
(i) Sometimes light behaves like a particle (called a photon), which explains how light
travels in straight lines
(ii) Sometimes it behaves like a wave, which explains how light bends (or diffracts)
around an object
The dual nature of electromagnetic radiation has been used to explain the atomic
and molecular processes of interaction of radiation with matter. The wave theory
postulates that radiation emanating from a source consist of an electromagnetic field
which varies periodically and in a direction perpendicular to the direction of propagation

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of radiation. The electromagnetic field is composed of electric and magnetic vectors

oscillating in mutually perpendicular plane. The radiation may be described in terms of a
sine wave based on periodic variations of the electric and magnetic field components.
Some important characteristics of wave
Every wave has five important characteristics, namely, wavelength (λ), frequency (υ),
velocity (c), wave number (ν–) and amplitude (a).
• Wavelength (λ): The distance between two neighboring troughs or crests is known as
wavelength. It is denoted by λ and is commonly expressed in nanometers (one nm = 10-9
m) or Angstrom (1 Å = 10-10 m).
• Frequency (υ): The frequency of a wave is the number of times a wave passes through
a given point in a medium per second. It is denoted by ν and is expressed in cycles per
second (cps) or hertz (Hz) 1Hz = 1cps. The frequency of a wave is inversely proportional
to its wavelength (λ).
υ ∝ 1/λ or υ= c/ λ
• Velocity(c): The distance travelled by the wave in one second is called its velocity. It is
denoted by c and is expressed in ms–1
c =υλ or λ = c/υ
• Wavenumber (ṽ): It is defined as number of wavelengths per cm. It is denoted by ṽ
and is expressed in cm–1
• Amplitude (a): It is the height of a crest or depth of a trough of a wave and is denoted
by a. It determines the intensity or brightness of the beam of light.
Photons
Light is made up of particles called photons. The energy of a photon is dependent
only upon its frequency.
E = hυ = hc/ λWhere h is Planck’s constant (h = 6.626 x 10-34 J-s).
The behavior of electromagnetic radiation depends on its wavelength or energy (i.e.,
frequency). Higher frequencies have shorter wavelengths and, vice versa, lower
frequencies haven longer wavelengths. The interaction of electromagnetic radiation with
atoms or molecules depends on the energy. Depending on the spectral region of interest,
the units used to describe the radiation may be frequency, energy, wavelength or inverse
wavelength (wavenumber).

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Reflection, Refraction and Diffraction

 Reflection involves a change in direction of waves when they bounce off a barrier.
 Refraction of waves involves a change in the direction of waves as they pass from
one medium to another. Refraction, or the bending of the path of the waves, is
accompanied by a change in speed and wavelength of the waves.
 Diffraction involves a change in direction of waves as they pass through an
opening or around a barrier in their path.

Energy Levels in a Molecule

In addition to the ordinary energy of transitional motion, a molecule may possess internal
energy, can be subdivided in to three classes: rotational energy, vibrational energy and
electronic energy.

(a) Translational energy (E trans): This energy is concerned with the translational
motion of molecules along three axes. It is significant only in gases and to a lesser extent
in liquids. In solids, molecules have fixed position, so generally solids are not associated
with translational energy.

(b) Rotational energy (E rot): This energy is associated with the spinning of molecules
about the axes passing through centre of gravity.

(c) Vibrational energy (E vib): This energy is associated with to and fro motion
(vibrations) of the molecule. Examples of such vibrations are stretching and bending.

(d) Electronic energy (E elec): Electronic energy is associated with the motion of
electrons. It deals with the transition of an electron from ground state to an excited state
due to the absorption of energy.

Total energy (E tot) of a molecule can be expressed as the sum of all these energies.

E tot = E elec + E vib + E rot + E trans

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Energy levels in atom

The electronic energy levels of atom are usually designated as S0 for the ground
state or lowest energy level and S1, S2, S3,…,etc., for the excited state having higher
energies. Atoms do not have vibrational or rotational energy level. The occupancy of the
electronic energy level in atom is determined by quantum theory and Maxwell-
Boltzmann distribution.

Where,

N upper and N lower – population of electron in the higher and lower energy levels
ΔE – energy separations between the levels
k – Boltzmann constant (1.3806*10-23 JK-1)
T – Absolute temperature
Based on these rule the ground state will be most occupied energy level and hence,
most of the atoms are said to be in their ground state under ambient conditions.
Energy level in molecules
The electronic energy levels of molecule are usually designated as S0 for the
ground state or lowest energy level and S1, S2, S3,…,etc., for the excited state having
higher energies. It also consist of sub energy level called vibrational energy levels
designated as V= 0,1,2,3 etc., Each vibrational energy level in turn consist of sub energy
levels called rotational energy levels designated as j = 0, 1,2,3, etc., The relative
magnitude of all energies are in the order of

E elec > E vib > E rot > E nuclear spin

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Majority of molecules exist in their ground state level of rotational, vibrational and
electronic energy levels. Molecules in addition to energy levels S1, S2, S3, (known as
singlet energy level) have also excited state called triplet energy levels designated as T1,
T2, etc. the lowest triplet energy level T1 has slightly lower energy compared to the S1
level. In excited singlet state the promoted electron, retain the original spin of anti-
parallel orientation to the electron in the ground state. However, in triplet state the
electrons occupying the ground and higher energy levels have parallel spins.
Interaction of photon with matter
When electromagnetic radiation passes through matter, a variety of phenomenon
may occur. Some of these are as follows:

 If the photon of radiation possess the appropriate energies, they may be absorbed
by the matter and result in electronic transitions, vibrational changes, rotational
changes or combinations of these. After absorption, atom and molecule become
excited. They give out energy quickly either by losing energy in the form of heat
or by re-emitting electromagnetic radiations.
 It is not necessary that the radiation passing through the matter may be absorbed
completely. The portion of electromagnetic radiation passes in to the matter,
instead of being absorbed, may undergo scattering or reflection or may be re-
emitted at the same or different wavelength.
 When electromagnetic radiation is neither absorbed nor scattered, it may undergo
changes, orientation, or polarization.
 In some cases, molecules after absorbing radiation become excited but they do not
lose energy quickly but with some delay. In such cases, energy is re-emitted as
radiation, usually of longer wavelength: than was originally absorbed. This
phenomenon is known as fluorescence.
 If in some cases, there is detectable time delay in re-emission, the phenomenon is
termed as phosphorescence.
Interaction of electromagnetic radiations with atom
 The interaction between electromagnetic radiation and atom may involve the
transfer of energy of incident photons to the atoms, which raises their energy from
the ground state to the excited state. This process is called as absorption denoted
as A.
 The absorption of energy by atoms results in the decrease in the intensity of
incident radiation at a particular frequency after it passes through the sample.

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 The intensity of the transmitted light is relatively weaker compared to the intensity
of incident radiation.
 The intensity of absorbance is proportional to the concentration of the absorbing
species.
 The excited atoms are stable for only for a short time.
 They return back to the ground state by radiative or non- radiative pathways
 The kinetic energy imparted to the atom is utilized to excite electrons to higher
energy level from which they relax to the ground state by emitting radiation. This
phenomenon is called as emission (E).
 In non – radiative pathway the energy is converted in to transitional energy and
evolves as heat
 The excited state atom may relax by radiative pathway referred to as fluorescence

Energy level diagram of an atom

Interaction of electromagnetic radiation with molecules

 Molecules also interact with electromagnetic radiations by absorbing energy from

the incident radiation. The absorption process in molecules can occur in different
regions of electromagnetic spectrum giving rise to molecular absorption
spectroscopy
 The excited state molecules can undergo relaxation to the ground state by emission
of radiation in the form of fluorescence (F) and phosphorescence (P).
 Fluorescence (F) emission always occur from the first excited electronic state to
any one of the vibrational energy level in the ground state
 Fluorescence (F) emission is quite intense as it is a spin-allowed transition.
 phosphorescence (P) is a spin forbidden transitions and hence, weak in intensity
the excited state molecule may relax to the ground state via non radiative
pathways such as vibrational relaxations (VR) and internal conversion (IC)

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 The process of non- radiative transfer from the singlet state to the lower energy
triplet state is called as intersystem crossing (ISC)
 The molecule in the vibrational excited state relaxes to the lowest vibrational level
of T1 by vibrational relaxation.
Absorbance
Absorbance is the measure of the capacity of a substance to absorb light of a
specified wavelength. It is equal to the logarithm of the reciprocal of the transmittance. It
is also known as optical density. The property is measured using spectroscopy,
particularly for quantitative analysis. Typical units of absorbance are called "absorbance
units," which have the abbreviation AU and are dimensionless.
Transmittance
Transmittance (T) is defined as the fraction of incident light which is transmitted,
ie, passes through, a sample. Thus, T = I/Io, where Io equals the intensity of light which
strikes the sample and I is the intensity of light after passing through the sample.
Relation between absorbance and transmittance
Beer – Lambert Law
When a visible light from a source falls on a colored homogeneous medium, a
portion of light is absorbed by the medium, a portion is reflected and the rest is
transmitted.

The intensity of incident light I0 is the sum of the intensities of absorbed (Ia), reflected
(Ir), and transmitted light (It).

The magnitude of Ir is quite small and can be neglected.

Lambert’s law states that when a monochromatic light passes through a transparent
medium, the rate of decrease in the intensity with the thickness of the medium is
proportional to the intensity of the incident light. The differential form of the law is given
as

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Where t is the thickness of the medium and k is the proportionality factor. The integral
form of above equation is given as

By changing from natural to common logarithm we get,

Beer investigated the effect of concentration of the absorbing medium on the intensity of
light and proposed the law that the intensity of transmitted beam of radiation passing
through an absorbing medium decreases exponentially as the concentration of absorbing
substance increases arithmetically. Beer’s law may be expressed as

Where k’ is another constant and c refers to the concentration of the absorbing substance.

Beer – lambert aw is the combined form of the above two expressions and is given as

Where A is the absorbance or optical density, ɛ is molar absorption coefficient or molar

absorptivity. Absorbance A is related to transmittance as
A= log (1/T) or (- log T)

A diagrammatic representation of Beer – Lambert equation

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Ultraviolet spectroscopy
The wavelength of range of UV radiation starts at the blue end of the visible light
about 4000Ȧ and ends at 2000 Ȧ. The ultraviolet region is subdivided in to two spectral
regions
 The region between 2000 Ȧ - 4000 Ȧ is knowns as near ultraviolet region
 The region below 2000 Ȧ is called the far or vacuum ultraviolet region

The UV radiation has sufficient energy to excite valence electrons in many atoms or
molecules; consequently, UV is involved with electronic excitation.
Excitation by UV and visible radiations
UV absorption spectra arise from transition of electrons within a molecule or an
ion from a lower to a higher electronic level and the UV emission spectra arise from the
reverse type of transitions. For a radiation to cause excitation, it must be in the UV region
of electromagnetic spectrum.
When a molecule absorbs UV radiation of frequency ν sec-1, the electron in that
molecule undergoes transition from lower to higher energy level. The energy difference
is given by
E=hν erg
The actual amount of energy required depends on the difference in energy between
the ground state E0 and excited state E1 nof the electrons.
E1- E0 = hν
Energy absorbed in UV region produces changes in the electronic energy of the molecule
resulting from transition of valence electrons in the molecules. Three distinct type of
electrons are involved in organic molecules. They are

 σ electrons – These electrons are involved in saturated bonds (eg., bond between
carbon and hydrogen in paraffin). As the amount of energy required to excite
electrons in these bonds is much more than that produced by UV light, compounds
containing σ bonds do not absorb UV radiations. Fr this reason, paraffin
compounds are frequently very useful as solvents
 π electrons – These electrons are involved in unsaturated hydrocarbons. Trienes
and aromatic compounds have these bonds.

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 N- electrons - these are electrons which are not involved in the bonding between
atoms in molecule. Examples are organic compounds containing nitrogen, oxygen
and halogen.
 As N- electrons can be excited by UV radiations, ant compounds that contain
atoms like nitrogen, oxygen, sulfur, halogen compounds or unsaturated
hydrocarbons may absorb UV radiations.
Electronic transitions in inorganic compounds effected by UV, and visible radiations
When molecules are electronically excited, the electron go from bonding to
antibonding orbital.

 Electrons in σ bond are excited to antibonding σ orbitals

 Electrons in π bond are excited to antibonding π orbitals
 n Electrons are excited to σ antibonding and π antibonding orbital
The change of electron density and energy necessary to cause excitation have
provided basis for the colorimetric analysis and pH indicators. Molecular group that
absorb UV or visible light are called as chromophore.

Structure of phenolphthalein in acid and basic solution

There is a change in the structure of molecule and a significant change in the
energy level of the chromophore. This results in the molecule absorbing visible radiation
when it is in alkaline solution and not in acid solution. Such changes are the basis of
many acid – base indicators.
Electronic transitions in organic compounds effected by UV, and visible radiations
Energy absorbed in the ultraviolet region by complex organic molecules cause
transitions of valence electrons in the molecules. These transitions are

 σ to σ*
 n to σ*
 n to π*

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 π to π*

Various energy level of a molecule Electronic excitation energies

The change in energy values for different transitions are in the order of

(a) n to π* Transitions
 These types of transition are shown by unsaturated molecule containing atoms
such as oxygen, nitrogen and Sulphur
 These transition exhibit a weak band in their absorption spectrum
 In aldehydes and ketones, the band occur in the range of 270-300 nm
 Carbonyl compounds having double bond separated by two or more single bonds
exhibit band in the range of 300-350 nm
 In aldehydes and ketones, transition arise from excitation of lone pair of electron
in 2py orbital of oxygen atom in to antibonding π* orbital of carbonyl compounds

Representation of n to π* transition
Methods to characterize n to π* transition
 One method is to observe the spectrum in the acid solutions. Bands due to these
transitions disappear in acidic solution due to a bond formed between the acidic
proton and lone pair of electron in heteroatom. Eg., band in pyridine disappears in
acidic solution

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 Another method to characterize the transition is done by comparing the compound

containing the hetero atom with the similar not containing heteroatom. The
spectrum of pyrazine exhibit band at 300 nm while spectrum of benzene doesn’t
exhibit these band.
(b) σ to σ* Transitions
 These transition occurs in compounds in which all the electrons are involved in
single bonds and there are no lone pair of electrons. Saturated hydrocarbons
involve in these transitions.
 As the energy required for these transitions are very large, the absorption band
occur in the far ultraviolet region (126-135 nm).
 methane at 121.9 nm and ethane at 135 nm corresponds to these transitions.
(c) n to σ* Transitions

 Saturated compounds with lone pair electrons undergo these transition along with
σ to σ* Transitions
 the energy required for these transition is less and absorption band appear at
longer wavelength in the near ultraviolet region (180-200 nm)
 In saturated alkyl halides the energy required for such transitions decreases with
increase in the size of halogen atom (or decrease in electronegativity of atom)
 The transition is more probable in methyl iodide as compared to methyl chloride
 Amines are known to absorb at higher wavelength as compared to alcohols and
hence the extinction coefficient for amine will be larger
 Hydrogen bonding shifts the ultra violet absorption to shorter wave lengths
(d) π to π* Transitions
 This transition corresponds to the promotion of an electron from a bonding π
orbital to antibonding π* orbitals.
 These transition can occur in any molecule having π electron system
 Selection rules determine whether a transition to a particular orbital is allowed or
not
 The spectrum of ethylene exhibits an intense band at 174 nm and weak band at
200 nm.
 According to selection rules the band at 174 nm is only allowed.
 The trans isomer absorbs at the longer wavelength with the greater intensity than
the cis isomer

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Summary of electronic structure and transition

Absorption wavelength of Typical organic functional groups

Choice of solvent
A suitable solvent for ultraviolet spectroscopy should meet the following
requirement

 It should not itself absorb radiations in the region under investigation

 It should be less polar so that it has minimum interaction with the solute molecules
The most commonly employed solvent is 95% ethanol. It is cheap, has good dissolving
power and doesn’t absorb radiation above 210 nm. In other words, it is transparent above

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210 nm. Commercial ethanol should not be used as it contains some benzene which
undergoes absorption in the UV range at 280 nm.
Some other solvent which are transparent above 210 nm are n-hexane, cyclohexane,
methanol, water and ether. Benzene, chloroform and carbon tetra chloride cannot be used
because they absorb in the range of about 240-280 nm.
Hexane and other hydrocarbons are sometimes preferred to polar solvents because they
have minimum interactions with the solute molecules.
The absorbance of solvents should be checked before use because the presence of small
amount of impurities may give rise to appreciable absorption in the range of sample.

List of common solvents generally used for recording UV spectra

Cut-off wavelength for solvents

The UV-Vis spectra are usually measured in very dilute solutions and the most
important criterion in the choice of solvent is that the solvent must be transparent within
the wavelength range being examined. some common solvents with their lower
wavelength cut off limits are listed below. Below these limits, the solvents show
excessive absorbance and should not be used to determine UV spectrum of a sample.
Highly pure, non-polar solvents such as saturated hydrocarbons do not interact
with solute molecules either in the ground or excited state and the absorption spectrum of
a compound in these solvents is similar to the one in a pure gaseous state. However, polar
solvents such as water, alcohols etc. may stabilize or destabilize the molecular orbitals of
a molecule either in the ground state or in excited state and the spectrum of a compound
in these solvents may significantly vary from the one recorded in a hydrocarbon solvent.

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Components of Optical Instruments

Components of Optical Instruments employed in optical spectroscopic methods
are based upon a) Absorption b) Scattering and c) Emission. Although the instruments for
measuring each differ somewhat in configuration, most of their basic components are
remarkably similar.
Components of typical spectroscopic instruments:

 A stable radiation source

 Wavelength selector
 Sample container (cell)
 A radiation detector
 A signal processor and readout

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Radiation Source
For proper measurements the radiation source must have following characteristics.

 It must generate a beam of sufficient power for ready detection and measurement.
 Source should provide continuous radiation in the region being studied.
 Source should be stable over long periods of time.
 Source should emit measurable signal throughout the region.
Sources of visible radiation: The most common visible radiation source is incandescent
tungsten filament lamp.
Sources of UV radiation: Hydrogen and deuterium lamps are most common sources
used in UV spectroscopy. Deuterium lamp produces continuous radiation of higher
intensity under same operating conditions when compared to hydrogen lamp.
Sources of infrared radiation: Nernst glower (a mixture of ZrO2, Y2O3 and Er2O3) and
globar are most widely used. Nichrome wire (near IR and IR) and Tungsten filament
(near IR) can also be used.
Source of electromagnetic radiation can also be classified according to whether they
produce continuous or non-continuous (line) spectra.
Continuous spectra:
Continuum sources emit radiation that changes in intensity only slowly as a
function of wavelength. Most of the sources are incandescent. Continuous sources are
widely used in absorption and fluorescence spectroscopy. Examples: Lamps of
hydrogen, deuterium etc.
Line spectra:
These are the sources that emit a discrete wavelength (few discrete lines) required
for some instrumental applications. They are widely used in atomic absorption, atomic
and molecular fluorescence, Raman, polarimetry and refractometer.
Monochromators
Monochromator is used to separate a polychromatic radiation into suitable
monochromatic form. Monochromator is a device that resolves a radiation into its
component wavelengths and permits the isolation of any desired portion of the spectrum.

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Monochromator has the following components

 Entrance slit: Slit is positioned on the way of radiation which deflects the beam
and admits beam to pass through. It provides rectangular optical image.
 Collimating lens or mirror: It makes light beams parallel.
 Dispersing device (Prism or grating): Most widely used dispersive devices are
prism or grating

Virtually all modern instruments incorporate grating in their monochromators because

gratings are cheaper and disperse radiation linearly as a function of wavelength.

 Focusing device: Again a lens or mirror is used for focusing the desired radiation
towards the exit slit.
 Exit slit: Isolate the wavelength band of interest. Three quantities determine the
merit of monochromator
 Effective bandwidth: It is the width of the transmittance band at half of
the transmission value.
 Peak percent transmittance: It shows percentage transmittance at the
central wavelength.
 Central wavelength: It is the wavelength corresponding to the center of
the effective bandwidth.
Types of Optical Instruments
Different optical instruments comes with different nomenclatures. Most of the
optical instruments have commons components like source, monochromator, detector etc.
as mentioned earlier. Common nomenclatures are given below.

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Spectroscope:
An optical instrument used for the visual identification of atomic emission lines.
Because atoms can absorb or emit radiation only at certain specific wavelengths defined
by electron transitions, the spectrum of each type of atom is directly related to its
structure. In its simplest form, a spectroscope is a viewing instrument consisting of a slit,
a collimator, a dispersing element, and a focusing objective.

 Slit: Entrance of the beam

 Collimator: A collimator is a special type of lens that "straightens out" light
coming in at various angles so that all of the light is travelling in the same
direction. The wave front is converted into a planar wave front.
 Dispersive element: Usually prism or grating
 Focusing objective: The focusing objective is just a lens system, such as that on a
telescope, that magnifies the spectrum and focuses it for viewing by eye

A spectroscope gives useful information, but it is only temporary. To capture

spectroscopic data permanently, the spectrograph was developed.
Spectrograph
A spectrograph operates on the same principles as a spectroscope, but it contains
some means to permanently capture an image of the spectrum. Early spectrographs
contained photographic cameras that captured the images on film that is continuously
exposed to the entire spectrum of dispersed radiation. Now photographic plates are
replaced by diode arrays using a camera or charge coupled device (CCD) cameras that
convert an optical signal into an electrical signal. They capture the image and transfer it
to video or computer for further analysis.
Spectrometer
A spectrometer is any instrument used to probe a property of light as a function of
its portion of the electromagnetic spectrum, typically its wavelength frequency, or
energy.

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Spectroscope Seeing The first instruments for

observing spectra were
visual, and they were
properly called
spectroscopes
Spectrograph Drawing (Recording) When spectra began to be
recorded on photographic
emulsion, just over a
century ago, the
instruments were called as
spectrograph. They
produced an image and
other instrument like
densitometers take
measurement from image
produced by measuring the
photographic density
Spectrometer Measuring When direct electronic
measurements came in to
replace the two step
process of recording and
measuring, the instruments
were called as spectrometer

Spectrophotometer
Spectrometer with a photo transducer (which converts light energy to electric
energy) is called Spectrophotometer. It reads relative intensities in the spectrum, rather
than the wavelengths of emission/absorption. With most spectrophotometers,

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CH8491 – Instrumental Methods of Analysis 2019-2020

measurements can be done in both ultraviolet and visible region of the spectrum, which is
not possible with colorimeters.

Spectrofluorometric or spectrofluorimeter: It is a spectrophotometer for fluorescence

analysis.
Colorimeter: Instruments used for absorption measurement using one or more color
comparison standards.

Photometer
A photometer is an instrument that measures intensity of photons. Photometers
measure absorption, scattering, reflection, fluorescence, phosphorescence etc. Photometer
consists of a source, filter, transducer, signal processor and read out. Photometer contains
a filter instead of dispersive device. Wavelengths can only be changed by changing the
filter. Some scientists refer photometers as colorimeters or photoelectric colorimeters.
Photometers used for fluorescence measurements are also called fluorometers.
Photons are quanta of electromagnetic energy that exhibit both wave-like and
particle-like properties. A photon can be considered to have a wavelength and frequency
(like a wave), as well as momentum and energy (like a particle). Despite carrying

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CH8491 – Instrumental Methods of Analysis 2019-2020

electromagnetic energy, a photon has no 'charge' and has a much lower chance of
interacting with matter than charged particles such as electrons and protons.
There are six ways in which photons may interact with matter:

 Coherent Scattering
 Photoelectric Effect
 Incoherent Scattering, also known as Compton Scattering or Compton
Effect
 Pair Production
 Triplet Production
 Photodisintegration
Coherent Scattering
Coherent (or Rayleigh) scattering occurs at low photon energies. A photon may
interact with an orbital electron and get deflected (or scattered) at a small angle. There is
no change in energy of the photon. This makes it of some concern in diagnostic x-ray,
where it can cause loss of contrast and blurring. For radiotherapy it has minimal impact
on attenuation.
Photoelectric Effect
The photoelectric effect occurs when a photon interacts with an orbital electron
whose binding energy is close to that of the photon energy. In this scenario, the photon
disappears and all of its energy is given to the orbital electron, which is then ejected from
the atom with kinetic energy equivalent to the photon energy minus the binding energy.
The space left by the departing electron is filled by another orbital electron, with
emission of characteristic x-rays or Auger electrons (when a characteristic x-ray causes
ejection of another orbital electron within the same atom). In soft tissues, these
characteristic x-rays have a low energy (about 500 eV) and are absorbed locally. The
probability of the photoelectric effect occurring is

 Inversely proportional to the cube of the photon energy

 Proportional the cube of the atomic number
Therefore, as photon energy increases, the likelihood of the photoelectric effect decreases
rapidly. As the atomic number of a material increases, the likelihood of the photoelectric
effect increases rapidly. The photoelectric effect ceases to become likely at energies over
100-200 keV in water, but remains an important factor for attenuation in lead at energies
up to 1 MeV.

St. Joseph’s College of Engineering Department of Chemical Engineering

CH8491 – Instrumental Methods of Analysis 2019-2020

Incoherent Scattering (Compton Effect)

Incoherent scattering is the most important interaction in radiotherapy. It occurs
when a photon has a much greater amount of energy than the binding energy of the
electron, effectively considering the electron as 'free'. In this interaction, the photon
interacts with the 'free' electron, giving up some of its energy and undergoing scattering.
The electron receives the energy and is set in motion in a different direction. Photons may
be scattered in any direction, whereas electrons are only scattered in a forward direction
(maximum 90o from the original path of the photon). The energy given to the electron is
dependent on the photon energy (low energies tend to give up minimal energy to the
electron) and the angle of impact.

 Direct impact scatters the photon at 180 degrees and gives a large amount
of energy to the electron which continuous in the photons original path.
 Glancing hits cause no photon scattering and give a small amount of energy
to the electron, which moves at 90 degrees away from the photon direction.
Importantly, incoherent scattering is not directly related to atomic number but rather the
concentration of electrons in tissue. Incoherent scattering also decreases with energy, but
nowhere near as rapidly as the photoelectric effect. This makes it more relevant as photon
energies rise above the K-shell binding energies of orbital electrons.
Pair Production - κ
Pair production occurs when a photon passes very close to the nucleus of an atom.
If the energy of the photon is high enough, the photon may disappear and 'create' an
electron and a positron. The new particles move away with the remaining energy of the
photon converted to kinetic energy. The positron will typically annihilate once it loses its
kinetic energy, giving rise to annihilation quanta (two photons with energy of 0.511 keV).
Pair production only occurs when photon energy is high enough and when there is an
object of sufficient mass to take on the momentum gained by the new particles.

 Photon energy must be at least 1.022 MeV to generate a positron/electron pair.

This is because 1.022 MeV is required to create the mass of the electron and the
positron (through E=mc2).
 Pair production only occurs in close proximity to the nucleus. This is so
momentum of the electron and positron is conserved.
It is possible for pair production to occur in proximity to an electron. This is called triplet
production. Pair production does not occur with photon energy under 1.022 MeV. Once

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this threshold is reached, it becomes more likely as photon energy increases. Pair
production is related to the atomic number of a material.

Triplet Production - κtr

Triplet production is a special case of pair production which occurs in the vicinity
of an orbital electron. The photon disappears and the energy is used to create an electron
and positron. The orbital electron also receives energy and is freed from the atom. The
threshold for this to occur is a photon of 2.044 MeV.
Photodisintegration – π
Photodisintegration is an uncommon event that occurs when a photon is absorbed
by the nucleus of an atom. The photon is destroyed and a nucleon (either a proton or a
neutron) is released. The threshold for this effect is over 10 MeV for most nuclei (with
the exception of beryllium and deuterium, where it is 2 MeV). Even at high energies,
photodisintegration is an uncommon event and does not attenuate a substantial portion of
a photon beam. It is more important for radiation protection concerns - if neutrons are
released they are highly penetrating and can convert atoms into unstable isotopes. The
release of a nucleon from the atom in question also usually results in a radioactive
daughter product. The production of neutrons in high energy linear accelerators means
that bunkers must be regularly ventilated to prevent buildup of radioactive gasses

St. Joseph’s College of Engineering Department of Chemical Engineering