Kerala Govt Polytechnic College Kozhikode Lecture note

Class- 1

INSTRUMENTATION AND PROCESS CONTROL

Spectroscopy

Spectroscopy deals with the study of interaction of electromagnetic radiation with matter. In
spectroscopic analysis, the atoms of a substance are excited, and they-either absorb or radiate
energy at unique frequencies. The absorption or emission of radiations are studied to identify
the components in a mixture.
Most organic molecules and functional groups are transparent in the portions of the
electromagnetic spectrum that we call the ultraviolet (UV) and visible (VIS) regions—that is,
the regions where wavelengths range from 190 nm to 800 nm.

When continuous radiation passes through a transparent material, a portion of the radiation
may be absorbed. If that occurs, the residual radiation, when it is passed through a prism, yields
a spectrum with gaps in it, called an absorption spectrum.
As a result of energy absorption, atoms or molecules pass from a state of low energy (the initial, or ground
state) to a state of higher energy (the excited state)
Kerala Govt Polytechnic College Kozhikode Lecture note

UV Visible spectrophotometer
In the case of ultraviolet and visible spectroscopy, the transitions that result in the absorption
of electromagnetic radiation in this region of the spectrum are transitions between electronic
energy levels. As a molecule absorbs energy, an electron is promoted from an occupied orbital
to an unoccupied orbital of greater potential energy.
Generally, the most probable transition is from the highest occupied molecular orbital (HOMO)
to the lowest unoccupied molecular orbital (LUMO). The energy differences between
electronic levels in most molecules vary from 125 to 650 kJ/mole (kilojoules per mole).

Beer–Lambert Law
A = log(I0/I ) = εcl for a given wavelength
A = absorbance
I0 = intensity of light incident upon sample cell
I = intensity of light leaving sample cell
c = molar concentration of solute
l = length of sample cell (cm)
Kerala Govt Polytechnic College Kozhikode Lecture note

ε= molar absorptivity

Ultraviolet absorption photometers operates in the range from 0.2 to 0.8 microns. An
image of the light source (tungsten lamp for use in the visible region from 0.3 to 1.0 microns
and a hydrogen-discharge lamp for use in the ultraviolet region from 0.22 to 0.35 micron) is
focused by the condensing mirror and by the flat mirror on the entrance slit. From the entrance
slit it proceeds to a collimating mirror and to a quartz prism. The quartz prism reflects the
radiations back to the collimating mirror and a portion of the spectrum passes through the exit
slit and then through the sample cell and finally falls on the phototube. The intensity of the
beam is measured by a suitable amplifier connected to an automatic balance potentiometer
instrument. The particular wavelength passed through the sample cell is determined by the
angle of setting of the prism. The angle of the quartz prism determines which part of the
refracted beam falls on the exit slit. The sample cell can be arranged for continuous flow of gas
and liquid samples.

Application of ultraviolet spectroscopy

Kerala Govt Polytechnic College Kozhikode Lecture note

The ultraviolet absorption spectroscopy in other respects is similar to infrared spectroscopy.

The instrument may be used either for complete analysis of samples by running a complete
spectrogram or for continuous analysis of process streams by operating at a fixed wavelength
critical to the analysis being made. They are useful in analysis of almost all substances. In
petroleum refining and synthetic rubber production they can be used for analysis of
hydrocarbons and gases. In pharmaceutical work, such as penicillin production, they are used
for determining structure. In chemical plants they are used for analysing both organic and
inorganic compounds.

Mass spectrometer and lists its application

Mass spectroscopy employs some of the most fundamental characteristics of materials.

a) For a given accelerating force, the acceleration is inversely proportional to the mass.
b) As the ions of various materials have different weights, they are accelerated at different
rates.
c) These ions follow a curved path when they are caused to pass through a magnetic field.
Kerala Govt Polytechnic College Kozhikode Lecture note

d) For a given voltage and magnetic field, the radius depends upon the mass.
In mass spectrometer the sample is first converted into ions, by bombarding it with a stream of
electrons. The application of a negatively charged field will then accelerate these particles and
as they are located in a magnetic field, they follow a curved path with a radius which is
proportional to the square root of the mass of the ion. Since each particle of different mass will
have a characteristic radius, it is possible to sort out each ion according to its mass. The value
of the current delivered by the ions is a measure of the degree of concentration of a particular
ion. By automatically varying the accelerating voltage, a spectrum of ion beam intensity vs ion
mass is recorded and the analysis of the spectrum results in the determination of the
composition of gas or vapour.
Important components of mass analysers are
i. Power supply
ii. Sample system
iii. High vacuum system
iv. Ionising and collecting systems and
v. Amplifying and recording system.
The power supply and control system provide the accelerating voltage and means for
controlling the strength of the magnetic field. The sampling system provides the samples
representative of the material being measured, at suitable rates. A high vacuum system is
necessary in order to remove the air and other gases which would invalidate the results. The
amplifiers are essential, which are used to amplify the very feeble currents (of the order of
10-10 amp.) created by the ions arriving at the collector plate, so that the conventional
recording or indicating equipment could be used.

Applications of mass spectrometer

The mass spectrometer is most useful in analysing aliphatic and cyclic hydrocarbon
compounds, such as those produced in aviation-gasoline and synthetic-rubber manufacture.
In petroleum refining, light gases, olefins, paraffin’s, wet gas, propylene, and etc. can be
analysed with mass spectrometer.
In chemical manufacturing, liquid and gaseous pharmaceuticals, organic compounds having
adequate vapour pressure, and inorganic gases and vapours may be analysed.

Disadvantages of mass spectroscopy

Kerala Govt Polytechnic College Kozhikode Lecture note

One of the major disadvantages is that it is difficult to distinguish between isomers of a

compound having the same charge-to-mass (m/z) ratio.

Chromatography

Chromatography is a technique of separation and purification of components

of a mixture by their differing affinities for two phases (states) of matter with
which they come into contact. Chromatography is based on the principle of
selective distribution of the different components of a mixture between two phases,
namely stationary phase and mobile (or moving) phase. The stationary phase can
be a solid or liquid, while the mobile phase is a liquid or gas. When the stationary
phase is solid, the selective distribution is based on adsorption and when it is a
liquid, the basis of selective distribution is partition.

Advantages of chromatography

i. It can be used for a mixture containing any number of components

ii. Very small quantities of the substances can be effectively detected and separated
from a mixture.
iii. Chromatography can be employed for purification of substances

iv. It may be used in some cases when other methods of analysing a mixture turn out to
be ineffective.
v. The method is simple and therefore it is widely used to analyse the most diverse
mixtures of inorganic and organic substances.

Adsorption chromatography

Adsorption (or column) chromatography is based on the selective adsorption of

different components of the mixture being analysed by using appropriate adsorbents.
The adsorbent must be finely divided to offer a vast surface area in proportion
to the particle size. On passing the mixed solution through the adsorbent column,
solutes are selectively adsorbed on the adsorbent particles with the help of ionic
bonds, hydrogen bonds or van der Waals forces.
Because of the different adsorbability and rate of movement of separate components,
in the solution being analysed, the different components of the mixture are held of
different height of the adsorption column, in the form of separate layers (or bands).
Kerala Govt Polytechnic College Kozhikode Lecture note

Components having great adsorbability are adsorbed in the upper part of the
adsorption column; while those having poorer adsorbability are situated lower.
Substances which are incapable of being adsorbed by a given adsorbent exist
through the column without being retained and accumulate in the filtrate.

In order to collect separately the adsorbed components, a proper solvent (or buffer
solution) is passed through the column so that individual adsorbed components of the
mixtures begin to emerge from the column which are collected. This process is
known as elusion. The process of elusion is continued until all the components of
the mixture have been collected in separate flasks. The samples collected are further
analysed to find out the composition.

Liquid column Chromatography

Liquid-Liquid Chromatography

A cylindrical column of an insoluble, inert and hydrophilic solid (like starch or

silica gel granules) is saturated with water, which remains immobilized around the
granules to form a stationary liquid phase. The mixture of solutes is dissolved in a
second immiscible solvent and the solution is passed through the column, the solvent
of this solution forming a mobile liquid phase. During the flow of the mobile
phase alongside the stationary liquid phase, solutes get partitioned (or distributed)
between the two liquid phases according to their partition coefficients, which
indicate their relative tendencies of dissolution in the two phases. This produces some
separation of the solutes from each other along the gel column. The separated solute
Kerala Govt Polytechnic College Kozhikode Lecture note

is then eluted with suitable solvent for further analysis.

Gas Solid Chromatography and Gas Liquid Chromatography

Gas chromatography is a technique by which a mixture is separated into its
constituents by a moving gas phase passing over a stationary adsorbent. The stationary
phase may be either liquid or solid. In this chromatography, if adsorption process is
taking place, the technique is known as gas-solid chromatography (GSC) ; while it
is called gas-liquid chromatography (GLC), when partition process is taking place.

Gas Solid Chromatography (GSC)

In GSC, the column is packed with an active solid stationary phase and the
components of the mixture are distributed between the gas phase and the active
solid phase. Due to differences in adsorption behaviour of components of mixture,
the separation is achieved. Common packing materials in GSC are - activated carbon,
silica gel or alumina.

Gas Liquid Chromatography (GLC)

In GLC, the column is packed with a porous solid (of large surface area), which is
coated with a thin-layer of a non-volatile liquid (stationary phase). Separation takes
place due to difference in partitioning of sample between the mobile gas phase
and thin-layer of liquid coated on inert support.
Common packing materials in GLC are porous material filled with non-volatile liquid
such as silicone oil, polyethylene glycol, grease, etc.

Theory (Common for GLC and GSC)

The sample mixture is introduced quickly into the moving carrier-gas (such as N2,
He, Ar, etc) The mixture is then swept along through the GLC or GSC column.
During the passage of mixture through column, the components of the mixture
distribute themselves
between the two phases. Some components are absorbed or adsorbed by the
stationary phase better than others, and hence, they are retained for longer time in
the column. Thus, each component of the mixture is carried along the column at
different rates and finally emerge from the column at different time. In other words,
Kerala Govt Polytechnic College Kozhikode Lecture note

the time for passage through the column is characteristic for each component. As
the component leaves the column, it is detected by instrumental means. A strip
chart recorder, attached to the detector, shows a peak for each component which is
separated. The size of the peaks indicate the amount of that particular component in
the mixture and its location on the chart indicates its nature.

The apparatus employed is called gas chromatograph and the series of peaks
obtained, is called chromatogram.

Comparison of GSC and GLC

Sl No Point of difference GSC GLC

1 Stationary phase Solid Liquid

2 Motile phase Gas Gas
3 Basis of separation Adsorption in the solid surface Partition between the gas
and liquid phase.
4 Packing of the column Adsorbents are packed in the Liquids are either coated as
form of fine size graded powder a fine film on the column
wall or coated as a fine film
on the inert support.
Kerala Govt Polytechnic College Kozhikode Lecture note

5 Length of column 0.7 to.2 m 3-300 m

6 Thermal stability of the Usually good. Only a few stationary phases

stationary phase are known to be stable
above 300°C
7 Reaction on the column Packing may catalyse to Vary rarely with the
produce some chemical change. stationary liquid, but may
occur due to interaction
with solid support
8 Usefulness Limited applicability, due to Vast applicability since
difficulty in reproducing surface large number of liquids of
conditions or due to surface Specific separation are
catalysis. available.

9 Applicability Useful in the separation of All volatile materials except

permanent gases and low boiling the more permanent gases.
substances.

High Pressure Liquid Chromatography ( HPLC )

In high pressure liquid chromatography, liquid rather than gas as the eluting
agent is used. The principle is much the same as in GLC. However, because
liquids are more viscous than gases, the pressure used to make them pass through
a column is greater than in GLC. The pressure is normally between 20 and 200 atm.
Such high pressures require a strong column, which is often about 25 cm in length.
The molecules coming of the column are detected by an ultraviolet
spectrophotometer, and the output appears as a series of peaks very much like
the GLC charts. HPLC is highly accurate and widely used in analysis and research.
Kerala Govt Polytechnic College Kozhikode Lecture note

Compare SEM and TEM

SEM TEM
Electron beam scans over surface of sample Beam passes through thin sample
Based on scattered electrons Based on transmitted electrons
Sample can ne of any thickness Thin samples
Samples mounted on aluminum stub Specimen supported on TEM grids
Image on monitor Image on fluorescent screen
Image is the surface of the sample Image is a two-dimensional projection of
sample
Shows image bit by bit Shows sample as a whole
Resolution 0.5 to 4 nanometer Resolution is 0.2nm