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EI6501 Analytical Instruments Dept.

of EIE and ICE 2017-2018

EI6501 -ANALYTICAL INSTRUMENTS


V SEMESTER (EIE & ICE) III YEAR PART B ANSWERS
UNIT I COLORIMETRY AND SPECTROPHOTOMETRY

1. Explain FTIR spectrophotometers in detail with necessary diagram. (Dec 2014, Dec 2015 Nov/Dec
2016, May/ June 2016)
The heart of FTIR spectrophotometer is a two beam interferometer most commonly of the
Michelson type.

Fourier Transform Infrared (FT-IR) spectrometry was developed in order to overcome the limitations
encountered with dispersive instruments. The main difficulty was the slow scanning process. A method
for measuring all of the infrared frequencies simultaneously, rather than individually, was needed. A
solution was developed which employed a very simple optical device called an interferometer. The
interferometer produces a unique type of signal which has all of the infrared frequencies encoded
into it. The signal can be measured very quickly, usually on the order of one second or so. Thus, the
time element per sample is reduced to a matter of a few seconds rather than several minutes.
Most interferometers employ a beam splitter which takes the incoming infrared beam and divides it
into two optical beams. One beam reflects off of a flat mirror which is fixed in place. The other beam
reflects off of a flat mirror which is on a mechanism which allows this mirror to move a very short
distance (typically a few millimeters) away from the beam splitter. The two beams reflect off of their
respective mirrors and are recombined when they meet back at the beam splitter. Because the path that
one beam travels is a fixed length and the other is constantly changing as its mirror moves, the signal
which exits the interferometer is the result of these two beams interfering with each other. The
resulting signal is called an interferograms which has the unique property that every data point (a
function of the moving mirror position) which makes up the signal has information about every
infrared frequency which comes from the source.
This means that as the interferograms is measured; all frequencies are being measured simultaneously.
Thus, the use of the interferometer results in extremely fast measurements. Because the analyst
requires a frequency spectrum (a plot of the intensity at each individual frequency) in order to make
identification, the measured interferograms signal cannot be interpreted directly. A means of
decoding the individual frequencies is required. This can be accomplished via a well-known
mathematical technique called the Fourier transformation. This transformation is performed by the
computer which then presents the user with the desired spectral information for analysis.

/
Some of the major advantages of FT-IR over the dispersive technique include: Speed: Because all of

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the frequencies are measured simultaneously, most measurements by FT-IR are made in a matter of
seconds rather than several minutes. This is sometimes referred to as the Felgett Advantage.
Sensitivity: Sensitivity is dramatically improved with FT-IR for many reasons. The detectors employed
are much more sensitive, the optical throughput is much higher (referred to as the Jacquinot
Advantage) which results in much lower noise levels, and the fast scans enable the coaddition of
several scans in order to reduce the random measurement noise to any desired level (referred to as
signal averaging). Mechanical Simplicity: The moving mirror in the interferometer is the only
continuously moving part in the instrument. Thus, there is very little possibility of mechanical
breakdown. Internally Calibrated: These instruments employ a HeNe laser as an internal wavelength
calibration standard (referred to as the Connes Advantage). These instruments are self-calibrating and
never need to be calibrated by the user.
2. Write short notes on: (Dec 2014)
(i) Attenuated total reflectance flame photometers (8)
Infrared methods can be used to study materials which are light scattering or opaque, or in the form of
coatings on opaque materials. The technique usually employed is based on obtaining of the reflectance
spectrum. However a spectrum obtained by reflection of the radiation from the surface of a chemical
material is generally very poor. This difficulty is overcome by Attenuated Total Reflectance which
enables one to obtain reflection spectra of satisfactory quality.
In this technique, energy from the source enters into a prism and is reflected almost entirely from face
C

the internal reflection will be attenuated or reduced at the sample-crystal surface. The attenuation will
take place at those wavelengths, where the material absorbs infrared energy. This facilitates
achievement of e absorption spectra of coatings and liquids even if they are opaque. The information
conveyed by the ATR spectra is essentially the same as that conveyed by the transmission spectra. The
optical arrangement of a simple ATR system shown in Figure.

(ii) Atomic absorption spectrophotometers. (8) (May 2013)


Atomic emission spectroscopy (AES) is a method of chemical analysis that uses the intensity of light
emitted from a flame, plasma, arc, or spark at a particular wavelength to determine the quantity of an
element in a sample. The wavelength of the atomic spectral line gives the identity of the element
while the intensity of the emitted light is proportional to the number of atoms of the element.
A sample of a material (analyte) is brought into the flame as a gas or sprayed solution. The heat from
the flame evaporates the solvent and breaks chemical bonds to create free atoms. The thermal energy
also excites the atoms into excited electronic states that subsequently emit light when they return to
the ground electronic state. Each element emits light at a characteristic wavelength, which is
dispersed by a grating or prism and detected in the spectrometer.
A frequent application of the emission measurement with the flame is the regulation of alkali metals

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EI6501 Analytical Instruments Dept. of EIE and ICE 2016-2017
for pharmaceutical analytics.
Diagram of Flame Emission Photometer:

3. What are the limitations of visual calorimeter? Explain the working principle of a double beam
calorimeter with a schematic diagram. (16) (May 2014)

limitations of visual calorimeter:


1 Heat loss through radiation and conduction.
2 Uniform temperature is not possible

In double beam fiber photometers, two photocells are normally employed. Two photocells are
connected to two phototmieter,P1 and P2 in Fig. each of the two potentiometers is of low resistance an
is would linearly. Light from the source lamp is made to pass through the filter F and is ten divided
into two parts. One part passing through the solution in the cuvette before falling on the measuring
photocell, and the other part passing directly on to the reference photocell. The galvanometer G
receives opposing currents through it.
4. With a block diagram, explain the operation of flame photometer. List the advantages of flame
photometry. (May 2014, May 2015, Nov/Dec 2016)
EI6501 Analytical Instruments Dept. of EIE and ICE 2016-2017

A flame photometer has three essential parts

5. State and derive Beers law from basic principles. Discuss the limitations of it.
A= bc
Where A is absorbance (no units, since A = log10 P0 / P
is the molar absorbtivity with units of L mol-1 cm-1
b is the path length of the sample - that is, the path length of the cuvette in which the sample is
contained. We will express this measurement in centimetres.
c is the concentration of the compound in solution, expressed in mol L-1

The linearity of the Beer-Lambert law is limited by chemical and instrumental factors. Causes
of nonlinearity include:

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1. deviations in absorptivity coefficients at high concentrations (>0.01M) due to


electrostatic interactions between molecules in close proximity
2. scattering of light due to particulates in the sample
3. fluorescence or phosphorescence of the sample
4. changes in refractive index at high analyte concentration
5. shifts in chemical equilibrium as a function of concentration
6. non-monochromatic radiation, deviations can be minimized by using a relatively flat part of
the absorption spectrum such as the maximum of an absorption band

6. With a schematic diagram explain the atomic emission spectroscopy.


Atomic emission spectroscopy (AES) is a method of chemical analysis that uses the intensity of light
emitted from a flame, plasma, arc, or spark at a particular wavelength to determine the quantity of an
element in a sample. The wavelength of the atomic spectral line gives the identity of the element while
the intensity of the emitted light is proportional to the number of atoms of the element.
A sample of a material (analyte) is brought into the flame as a gas or sprayed solution. The heat from
the flame evaporates the solvent and breaks chemical bonds to create free atoms. The thermal energy
also excites the atoms into excited electronic states that subsequently emit light when they return to the
ground electronic state. Each element emits light at a characteristic wavelength, which is dispersed by
a grating or prism and detected in the spectrometer.
A frequent application of the emission measurement with the flame is the regulation of alkali metals
for pharmaceutical analytics

7. Explain mono-chromators and detectors.

A monochromator is an optical device that transmits a mechanically selectable narrow band of


wavelengths of light or other radiation chosen from a wider range of wavelengths available at the input
A device that can produce monochromatic light has many uses in science and in optics because many
optical characteristics of a material are dependent on color. Although there are a number of useful
ways to produce pure colors, there are not as many other ways to easily select any pure color from a
wide range.
Prism may be made of glass or quartz. The glass prism may be suitable for radiations essentially in the
visible range whereas the quartz prism can over the ultraviolet spectrum also.

DETECTORS:
1.PHOTOVOLTAI CELL:

Construction:
This cell is also known as barrier layer cell or photronic cell. This cell is entirely different in design
and working principle from phototubes or photo emissive tube. There is no need of battery for the
operation of photovoltaic cell.
It usually consists of a metal base plate which may be iron or aluminum and which acts as one
electrode. A thin layer of semiconducting substance which is generally selenium deposited on the
surface of metal base plate. The semi conducting substance is covered with a thin layer of silver or
gold which acts as another collector electrode.
Operation:
When the radiation is incident upon the surface of the selenium through the transparent glass, it excites
the electrons at silver-selenium interface. The released electrons are collected by the silver. The
accumulation of more number of electrons in the silver surface creates potential difference between the
two electrodes. The potential difference creates a current flow. This current flow could be measured by
connecting a galvanometer between two electrodes. The current thus measured gives the amount of
radiation that falls on the photovoltaic cell

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1. PHOTOEMISSIVE TUBE:

When a beam of light falls on the surface of the cathode, electrons are released from it, which are
accelerated towards the anode which is maintained at certain positive potential by the battery. The
current generated by the radiation of light will not be sufficient to drive the read out devices. So it can
be amplified by an amplifier and the output current can be read by connecting a galvanometer after
the amplifier circuit. This is the measure of incident radiation.

8. Explain about photo multiplier tube.


Schematic Diagram:
A photomultiplier tube, useful for light detection of very weak signals, is a photoemissive device in
which the absorption of a photon results in the emission of an electron. These detectors work by
amplifying the electrons generated by a photocathode exposed to a photon flux.

Photomultipliers acquire light through a glass or quartz window that covers a photosensitive surface,
called a photocathode, which then releases electrons that are multiplied by electrodes known as metal
channel dynodes. At the end of the dynode chain is an anode or collection electrode. Over a very large
range, the current flowing from the anode to ground is directly proportional to the photoelectron flux
generated by the photocathode.

The spectral response, quantum efficiency, sensitivity, and dark current of a photomultiplier tube are

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determined by the composition of the photocathode. The best photocathodes capable of responding to
visible light are less than 30 percent quantum efficient, meaning that 70 percent of the photons
impacting on the photocathode do not produce a photoelectron and are therefore not detected.
Photocathode thickness is an important variable that must be monitored to ensure the proper response
from absorbed photons. If the photocathode is too thick, more photons will be absorbed but fewer
electrons will be emitted from the back surface, but if it is too thin, too many photons will pass through
without being absorbed.

Electrons emitted by the photocathode are accelerated toward the dynode chain, which may contain up
to 14 elements. Focusing electrodes are usually present to ensure that photoelectrons emitted near the
edges of the photocathode will be likely to land on the first dynode. Upon impacting the first dynode, a
photoelectron will invoke the release of additional electron that are accelerated toward the next
dynode, and so on. The surface composition and geometry of the dynodes determines their ability to
serve as electron multipliers. Because gain varies with the voltage across the dynodes and the total
number of dynodes, electron gains of 10 are possible if 12-14 dynode stages are employed.

Photomultipliers produce a signal even in the absence of light due to dark current arising from thermal
emissions of electrons from the photocathode, leakage current between dynodes, as well as stray high-
energy radiation. Electronic noise also contributes to the dark current and is often included in the dark-
current value.
9. Explain different radiation source in absorption spectrophotometry.
Blackbody Sources:A hot material, such as an electrically heated filament in a light bulb, emits a
continuum spectrum of light. The spectrum is approximated by Planck's radiation law for blackbody
radiations: B = 121n13/c2)(1/exp (hrr/kT)- 1), Planck's radiation where /I is Planck's constant, v is
frequency, c is the speed of light, k is the Boltzmann constant, and T is temperature in k. The most
common incandescent lamps and their wavelength ranges are: Tungsten filament lamps: 350 mu - 2.5
um Clowbar: I - 40 um Nernst glower: 400 nm - 20 pm. Tungsten lamps are used in visible and near-
infrared (NIR) absorption spectroscopy, while the glowbar and Nernst glower are used for infrared
spectroscopy. This lamp consists of a tungsten filament enclosed in a glass envelope. It is cheap,
intense and reliable. A major portion of the energy emitted by a tungsten lamp is in the visible region
and only about 15 to 20 per cent is in the infrared region. For work in the ultraviolet (UV) region, a
hydrogen or deuterium discharge lamp is used. In these lamps, the envelope material of the lamp puts a
limit on the smallest wavelength, which can be transmitted. For example, quartz is suitable only up to
200 rim and fused silica up to 185 nm. The radiation from the discharge lamps is concentrated into
narrow wavelength regions of emission lines. Practically, there is no emission beyond 900 nm in these
lamps. For this reason, spectrophotometers for both the visible and ultraviolet regions always have two
light sources, which can be manually selected for an appropriate wavelength.
Discharge Lamps: Discharge lamps such as neon signs pass an electric current through a rare gas or
metal vapour to produce light. The electrons collide with gas atoms, exciting them to higher energy
levels which then decay to lower levels by emitting light. Low pressure lamps have a sharp line
emission characteristic of the atoms in the lamp, and high pressure lamps have broadened lines
superimposed on a continuum spectrum. The common discharge lamps and their wavelength ranges
are: Hydrogen or deuterium: 160 - 360 nm Mercury: 253.7 nm, and weaker lines in the near-UV and
visible Ne, Ar, Kr, Xe discharge lamps: many sharp lines throughout the near-UV to near-1R Xenon
arc: 300 - 1.3nm. Deuterium lamps are presently used as the UV source in UV-vis absorption
spectrophotometers. The sharp lines of the mercury and rare gas discharge lamps are useful for
wavelength calibration of optical instrumentation. For fluorescent work, an intense beam of ultraviolet
light is required. This requirement is met by a xenon arc or a mercury vapour lamp. A cooling
arrangement is absolutely necessary when these types of lamps are used. Mercury lamps are usually
run direct from the ac power line via a series ballast choke. This method gives some inherent lamp
power stabilization and automatically provides the necessary ionizing voltage. The ballast choke is
physically small and a fast warm-up to the lamp operating temperature is obtained. A deuterium arc
lamp provides emission of high intensity and adequate continuity in the 190-380 nm range. A quartz or

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silica envelope is necessary not only to provide a heat shield, but also to transmit the shorter
wavelengths of the ultraviolet radiation. The limiting factor is normally the lower limit of atmospheric
transmission at about 190 nm.
Lasers The term 'laser' has been coined by taking the first letters of the expression 'Light amplification
by simulated emission of radiation'. Although it is an amplifier, as suggested by the abbreviation, the
laser is invariably used as a generator of light. But its light is quite unlike the output of conventional
sources of light. The laser beam has spatial and temporal coherence, and is monochromatic (pure
wavelength). The beam is highly directional and exhibits high density energy which can be finely
focused. The laser is acquiring an increasingly important role in analytical chemistry and it is
important for the practicing analyst to understand both its capabilities and its .Lasers have a number of
advantages over conventional sources such as glow bars, continuous discharges, pulsed discharges and
X-ray tubes, which make them useful for analytical chemistry applications. These advantages include
higher intensity, monochromaticity, low beam divergence, the availability of short and ultra-short
pulses for studies of transient phenomena, and coherence (well-defined phase). Lasers have been used
in many spectroscopic applications such as Raman Spectroscopy. However, there is little that the laser
can offer for UVvisible spectrophotometry. Perhaps, a high peak power will allow a measurable
amount of light to get through an optically dense sample so that high absorbance can be measured.

10 Explain the working principle of various radiation sources and detectors in IR


Spectro-photometers. (Nov/Dec 2016)
Three common practical infrared sources are;
1. Globar rod
A Globar is used as thermal light source for infrared spectroscopy. It is a silicon carbide rod of 5
to 10 mm width and 20 to 50 mm length which is electrically heated up to 1,000 to 1,650 C
(1,830 to 3,000 F).

2. Nernst Filament.
it is in the form of a cylindrical rod or tube composed of a mixture of certain oxides such as
zirconium oxide (ZrO2), yttrium oxide (Y2O3) and erbium oxide (Er2O3) at a ratio of 90:7:3 by
weight. They operated by being electrically heated to about 2000 C. Initially they required
external heating because the material is an insulator at room temperature.

3. Nichrome wire.
Tightly wound spiral of Nichrome wire heated to 1100 K by electric current.

11 With a neat sketch, explain the instrumentation setup and working principle of IR spectrometer.
(May 2013)
IR SPECTROPHOTOMETERS:

The light our eyes see is but a small part of a broad spectrum of electromagnetic radiation. On the
immediate high energy side of the visible spectrum lies the ultraviolet, and on the low energy side is
the infrared. The portion of the infrared region most useful for analysis of organic compounds is not
immediately adjacent to the visible spectrum, but is that having a wavelength range from 2,500 to
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16,000 nm, with a corresponding frequency range from 1.9*1013 to 1.2*1014 Hz. Photon energies
associated with this part of the infrared (from 1 to 15 kcal/mole) are not large enough to excite
electrons, but may induce vibrational excitation of covalently bonded atoms and groups. The covalent
bonds in molecules are not rigid sticks or rods, such as found in molecular model kits, but are more
like stiff springs that can be stretched and bent.. We must now recognize that, in addition to the facile
rotation of groups about single bonds, molecules experience a wide variety of vibrational motions,
characteristic of their component atoms. Consequently, virtually all organic compounds will absorb
infrared radiation that corresponds in energy to these vibrations. Infrared spectrometers, similar in
principle to the UV-Visible spectrometer, permit chemists to obtain absorption spectra of compounds
that are a unique reflection of their molecular structure. An example of such a spectrum is that of the
flavouring agent vanillin, shown above.

12 Draw and Explain the schematic diagram of a typical double beam spectrophotometer. (Nov
2013)
Schematic Diagram of Double Beam Spectrometer:

Working Principle:
The light beam from the source is split into sample beam and reference beam by the mechanical
chopper. The reference beam monitors the lamp energy whereas the sample beam reflects sample
absorption. The observed absorbance measurement is the ratio of the sample and reference beams
which are recombined before moving to the monochromator. This arrangement compensates the effects
due to drift in lamp intensity, electronic and mechanical fluctuations which affect both the sample and
reference beams equally.

13 What are non-dispersive spectrophotometers? Explain in detail the FTIR spectrometer. Also
specify the advantages of the FTIR spectrophotometer. (Nov 2013)

Non-dispersive spectrophotometers:
It is called non-dispersive because the wavelength which passes through the sampling chamber is not
pre-filtered, and instead the optical filter is in front of the detector to eliminate all light except the
wavelength which the selected gas molecules can absorb.
Advantages of FTIR are:-
1. FTIR stands for Fourier Transform Infrared Spectroscopy is highly sensitive and quick method
to achieve high quality spectrum.
2. This spectroscopy gives better signal to noise ratio compared to the dispersive instrument.
3. With FTIR, spectrum can be obtained very quickly and saves time.
4. Gases, solids as well as liquid can be analysed with FTIR.
5. By using FTIR no external calibration is required and gives accurate results.
6. FTIR is non-destructive technique.
7. Organic compounds and Inorganic compounds can be identified easily using fourier transform
infrared spectroscopy.
8. Mechanical breakdown is less compared to other instruments because the mirror which is attached
in the interferometer is the only moving part in FTIR.
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9. Simultaneous analysis can be done for multiple gaseous compounds.


10. FTIR can identify even small concentrations of contaminants.
11. FTIR has a laser beam which keeps the FTIR instrument accurately calibrated.
12. It generally completes a scans within 1 to 2 seconds.
13. High resolution.

14 Explain single and double beam instruments in detail. (Nov/Dec 2016)


Single beam instruments
Direct measurement of output of a single barrier-layer cell.
Suitable for a single beam instrument, which contains only one set of
optics.
Photoelectric current generated in the photocell by the light is given to
the galvanometer through the coarse and fine sensitivity controls at B
and A.
Double beam instruments
Provision is made for a second light path, which is emitted by an internal
standard element.
Internal standard and unknown wavelengths are isolated by means of optical
filters.
Photoelectric currents produced by the emitted radiations of unknown and
internal standard elements oppose each other through a galvanometer.
By means of accurate potentiometer P1, the opposing photoelectric currents
can be balanced.
Sensitivity is adjusted with another potentiometer P2, by shunting currents
generated by the measuring cell. This makes it possible to read the
concentration of the test solution without changing the concentration of
internal standard solution

Circuit diagram for double beam instruments


Circuit diagram for single beam instruments

15 Write a short note on fluorescent photometer. (May/ June 2016)


The basic instrument is a spectrofluorometer.
It contains a light source, two monochromators, a sample holder and a detector.
There are two monochromators, one for selection of the excitation wavelength, another for analysis
of the emitted light. The detector is at 90 degrees to the excitation beam.
Upon excitation of the sample molecules, the fluorescence is emitted in all directions and is detected
by photocell at right angles to the excitation light beam.

The lamp source used is a xenon arc lamp that emits radiation in the UV, visible and near-
infrared regions.
The light is directed by an optical system to the excitation monochromator, which allows
either pre selection of wavelength or scanning of certain wavelength range.
The exciting light then passes into the sample chamber which contains fluorescence cuvette. A special
fluorescent cuvette with four translucent quartz or glass sides is used
When the excited light impinges on the sample cell, molecules in the solution are excited
and some will emit light.
Light emitted at right angles to the incoming beam is analyzed by the emission
monochromator.
The wavelength analysis of emitted light is carried out by measuring the intensity of
fluorescence at preselected wavelength.
The analyzer monochromator directs emitted light of the preselected wavelength to the
detector.A photomultiplier tube serves as the detector to measure the intensity of the light.
The output current from the photomultiplier is fed to some measuring device that indicates the extent
of fluorescence.
UNIT II CHROMATOGRAPHY

1. Explain high pressure chromatographs in detail (16) (Dec 2014, May 2015, Dec 2015, Nov/Dec
2016, May 2016)

Applications:

Ideally suited for separation and identification of amino acids, proteins, nucleic acids,
hydrocarbons, carbohydrates, pharmaceuticals, pesticides, pigments, antibiotics, steroids, and a
variety of other inorganic substances.
Advantages to HPLC
Advantages of HPLC are result of 2 major advances:
stationary supports with very small particle sizes and large surface areas
appliance of high pressure to solvent flow

2. Explain with sketches the operation of any two detectors used in gas chromatography. (16) (Dec
2014)
Gas chromatography Detectors:
1. Flame ionization detector.
2. Photo ionization detector.

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MECHANISM: Compounds are burned in a hydrogen air flame. Carbon containing compounds
produce ions that are attracted to the collector. The number of ions hitting the collector is measured
and a signal is generated.

SELECTIVITY: compounds with C_H bonds. A poor response for some non-hydrgen containing
organics (eg: hexachloro benzene)

LINEAR RANGE: 105-107


TEMPERATURE: 250-300 Deg C

PHOTO IONIZATION DETECTOR (PID):

MECHANISM: Compounds eluting into a cell are bombarded with high energy photons emitted from
a lamp. Compounds with ionization potentials below the photon energy are ionized. The resulting ions
are attracted to an electrode, measured and a signal is generated.
LINEAR RANGE: 105-106
TEMPERATURE: 250-300 Deg C
3. Give in detail the classification of Chromatography. Briefly explain liquid chromatography. (16)
(May 2014).Chromatographic methods can be differentiated based on the physical means of bringing
the stationary and mobile phases into contact

. Column Chromatography - the stationary phase is held in a narrow tube through which the mobile
phase is forced either by pressure or by gravity. Examples include:

Simple column chromatography

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High pressure liquid chromatography (HPLC)
Gas chromatography (GC).

Column chromatography can be further differentiated based on the types of stationary and mobile
phases and the kinds of equilibria involved in solute transfer between the phases.

There are two broad categories in this classification scheme:

Liquid chromatography (for instance simple column or HPLC)


Gas Chromatography (GC).

2. Planar Chromatography - the stationary phase is supported on a flat plate or in the fibres of a
paper. Here the mobile phase moves through the stationary phase by capillary actionorbygravity.
Examples include:Paper chromatography

Thin layer chromatography (TLC).

General Specific
Stationary Phase Type of Equilibrium
Classification Method Name
Liquid Liquid-liquid Liquid adsorbed on Partition between
Chromatography or partition a solid immiscible liquids
(LC)
Mobile Phase = Liquid-bonded Organic species Partition between liquid
liquid phase bonded to a solid and a bonded surface
surface
Liquid-solid or Solid Adsorption
adsorption
Ion exchange Ion-exchange resin Ion exchange
Size exclusion Liquid in interstices Partition/sieving
of a polymeric solid
Gas Gas-liquid Liquid adsorbed on Partition between gas and
Chromatography a solid liquid
(GC)
Mobile Phase = gas Gas-bonded Organic species Partition between liquid
phase bonded to a solid and bonded surface
surface
Gas-solid Solid Adsorption

LIQUID CHROMATOGRAPHY:

Liquid chromatography is a technique used to separate a sample into its individual parts. This
separation occurs based on the interactions of the sample with the mobile and stationary phases.
Because there are many stationary/mobile phase combinations that can be employed when separating
a mixture, there are several different types of chromatography that are classified based on the
physical states of those phases. Liquid-solid column chromatography, the most popular
chromatography technique and the one discussed here, features a liquid mobile phase which slowly
filters down through the solid stationary phase, bringing the separated components with it.
4. With block diagram, explain the Gas chromatography. State the factors to be considered in
Carrier Gas supply system.(16) (May 2014 , May 2013, Nov 2013, Nov 2016, May 2016)

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Factors to be considered in Carrier Gas supply system:


1. The carrier gas must be chemically inert. That is otherwise it should not react with the sample or
stationary phase.
2. It should be suitable for the detector employed and the type of sample analysed.
3. It should give best column performance consistent with required speed of the analysis.
4. It should be free from the risk of fire.

5. Draw the schematic diagram of a gas chromatography and explain the different parts in Gas
chromatography.(May 2013), (Nov 2013)

The basic parts of GC are


1. Carrier gas supply along with pressure regulator and flow meter
2. Sample injection system
3. Chromatographic column
4. Thermal compartment or thermostat
5. The detection system
6. Recorder

CARRIER GAS SUPPLY ALONG WITH PRESSURE REGULATOR AND FLOW METER
The carrier gas normally N2, Ar, or He , compressed in cylinder with suitable pressure regulator.
The sample injection port is maintained at temp T1, which ensures rapid vaporization but not
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thermal degradation of the solute.
The gas and liquid sample are injected by syringe through a self-sealing silicon rubber diaphragm
in the injection port.
SAMPLE INJCTION SYSTEM
For optimum column efficiency, the sample should not be too large, and should be introduced onto
the column as a "plug" of vapour - slow injection of large samples causes band broadening and
loss of resolution.
The most common injection method is where a microsyringe is used to inject sample through a
rubber septum into a flash vapouriser port at the head of the column. The temperature of the
sample port is usually about 50C higher than the boiling point of the least volatile component of
the sample.

CHROMATOGRAPHIC COLUMN
The column is the heart of a gas chromatograph wherein the fundamental process of separation
takes place.
There are two general types of column, packed and capillary (also known as open tubular).
Packed columns contain a finely divided, inert, solid support material (commonly based on
diatomaceous earth) coated with liquid stationary phase. Most packed columns are 1 and 50m in
length and have an internal diameter of 4 - 8mm.
Packed columns are made from Glass, stainless steel or copper.

THERMAL COMPARTMENT
1. The column is not operated at room temp because it would then be suitable only for the
analysis of gases or extremely volatile liquids.
2. It is desirable to keep the column at precisely constant temp. This is essential because
the quantitative response of the detector is often affected by column temp
3. The temp is controlled to an accuracy of 0.1deg C.
4. The temp of the oven can be controlled accurately by using a proportional temp controller with
a platinum resistance thermometer as sensing element.
5. THE DETECTION SYSTEM AND RECORDER
6. Convert the sample into electrical signal and record the signal as chromatogram.

6. Describe a gas liquid chromatography with a schematic diagram. Explain how it


works. Schematic Diagram of Gas-liquid Chromatography: (May 2016)

The mobile liquid phase is made to percolate through a column containing the stationary phase
which deposited on a solid surface as a thin film.
The solid surfaces used are silica gel, porous glass and cellulose.

7. Give a typical chromatogram. Discuss some applications of gas liquid chromatography.

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Typical chromatogram would comprises a series of peaks separated in the domain, each peak
corresponding a chemical component detected.
The area enclosed by the peak corresponds to the concentration of that chemical.
The area of the peak is measured by counting squares using planimeter or cutting out the peak
and weighing the paper.
Width at 50% heightheight:
The peak height(h) is measured and multiplied by the width at half-
height(w) Area = w.h
Triangulation: Drawing tangents to each side of the peak and to extend them so as to form a
triangle with base line. The area of this triangle is approximated proportional to the area under the
peak. Area of the triangle = 1/2.w.h, w- peak width & h-peak height.
Weighing of the chart paper
The peak area is determined by cutting out the peak from the chart and weighing the paper. The
method assumes a uniform weight distribution in paper so that the area is proportional to weight.

8. Explain in detail the following detectors used in Gas Chromatography.


a).Thermal conductivity detector b).Flame ionization detector

Thermal conductivity detector


1. The TCD consists of an electrically heated filament in a temperature-controlled cell. Under
normal conditions there is a stable heat flow from the filament to the detector body
2. .
3. When an analyte elutes and the thermal conductivity of the column effluent is reduced, the
filament heats up and changes resistance.

4. This resistance change is often sensed by a Wheatstone bridge circuit which produces a
measurable voltage change. The column effluent flows over one of the resistors while
the reference flow is over a second resistor in the four-resistor circuit.

5. Since all compounds, organic and inorganic, have a thermal conductivity different from
helium, all compounds can be detected by this detector.

6.The TCD is often called a universal detector because it responds to all compounds. Also, since
the thermal conductivity of organic compounds is similar and very different from helium, a
TCD will respond similarly to similar concentrations of analyte.
Schematic Diagram:

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FLAME IONISATION DETECTOR:


An FID consists of a hydrogen/air flame and a collector plate. The effluent from the GC
column passes through the flame, which breaks down organic molecules and produces ions.
The ions are collected on a biased electrode and produce an electrical signal. The FID is
extremely sensitive with a large dynamic range; its only disadvantage is that it destroys the
sample.
It responds with high sensitivity to almost all organic compounds.

9. With a neat schematic diagram discuss the separation principle of HPLC. (May 2013)
The most widely used analytical separations technique
Utilizes a liquid mobile phase to separate components of mixture
uses high pressure to push solvent through the column
Popularity:
sensitivity
ready adaptability to accurate quantitative determination
suitability for separating nonvolatile species or thermally fragile ones
widespread applicability to substances that are of prime interest to industry, to many
fields of science, and to the public
Ideally suited for separation and identification of amino acids, proteins, nucleic acids,
hydrocarbons, carbohydrates, pharmaceuticals, pesticides, pigments, antibiotics, steroids, and a
variety of other inorganic substances
Advantages of HPLC are result of 2 major advances:
stationary supports with very small particle sizes and large surface areas
appliance of high pressure to solvent flow

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10. What are the requirements of HPLC pumping system and enumerate the application of HPLC.
Pumping System I
Provide a continuous constant flow of the solvent through the injector
Requirements
pressure outputs up to 6000 psi
pulse-free output
flow rates ranging from .1-10 mL/min
flow control and flow reproducibility of .5% or better
corrosion-resistant components
SINGLE PISTON-RECIPROCATING PUMP
On the back stroke the separation column valve is closed and the piston pulls in solvent from
the mobile phase reservoir.
On the forward stroke the pump pushes solvent out to the column from the reservoir.
Range of flow rates can be attained by altering the piston stroke volume during each cycle or
by altering the stroke frequency.
Pumping System II
Two types:
constant-pressure
constant-flow
Reciprocating pumps
motor-driven piston
disadvantage: pulsed flow creates noise
advantages: small internal volume (35-
ready adaptability to gradient elution, constant flow rates
Pumping System III
Syringe or Displacement pumps
syringe-like chambers activated by screw-driven mechanism powered by a stepper motor

advantages: output is pulse free


disadvantage: limited solvent capacity (~20 mL) and inconvenience when solvents need
to be changed
Flow control and programming system
computer-controlled devices
measure flow rate
increase/decrease speed of pump motor
APPLICATION OF HPLC
1. HPLC is applied in the process of isolation and purification of compounds
2. Chemical separation can be accomplished using HPLC by utilizing the fact that certain
compounds have different migration rates with respect to the particular column and mobile
phase.
3. It is also applied for the purification process that is to separate or extract the target compound
from other compounds to contaminants
4. A particular compound could be identified with greater accuracy.

11. Explain various detectors used in liquid chromatography.


1. Mercury or xenon source.
2. Conductivity Detector.
3. Fluorescence Detector
12. Explain about Refractive index detector and electrochemical detector
1. Refractive index detector

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Working Principle:
Based on the principle of refraction, reflection or interference of light beams.
Based on the means of difference in RI between the solution and of the pure solvent.
Cell is divided into 2 chambers by a glass plate which is mounted at an angle such that bending of
the incident beam occurs if the 2 solutions differ in RI.
An optical mask is placed in front of the lamp to confine the beam of light.
The beam collimated by a lens passes through both compartment and is reflected back by the
mirror through the compartment again.
If the RI of the mobile phase is changed due to presence of the solute, the beam from the sample
compartment is slightly deflected.
As the beam changes location on the detector an out of balance signal is generated in the detector
and that is proportional to the concentration of the solute.
An optical flat is used to adjust for zero output signals when both the compartment contains the
pure mobile phase.
2. ELECTROCHEMICAL DETECTOR:

Schematic Diagram:

Working Principle:
Based on reduction or oxidation of the eluting compound at a suitable electrode and measurement
of resulting current
Amperometric transducer measures the Nano ampere level current.
Teflon gasket divides the chamber into 2 channels.
One side of the channel working electrode is positioned. Reference electrode and often a counter
electrode are located in downstream from the working or indicator electrode.
The eluent first flows over one electrode (upstream) and then over the second electrode
(downstream).
The upstream working electrode generates an electroactive product from the analyte.
That product is subsequently detected at the downstream working electrode.
Advantages
high sensitivity
ease of use

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Disadvantages
mobile phase must be made conductive
Mobile phase must be purified from oxygen, metal contamination, halides.
13. Explain with neat diagram about flame photometric detector, photo ionization detector and
electron capture detector.
1.Flame photometric detector:
Schematic Diagram:

The instrumental requirements for 1) a combustion chamber to house the flame, 2) gas lines for
hydrogen (fuel) and air (oxidant), and 3) an exhaust chimney to remove combustion products, the
final component necessary for this instrument is a thermal (band pass) filter to isolate only the
visible and UV radiation emitted by the flame.
Without this the large amounts of infrared radiation emitted by the flame's combustion reaction
would heat up the PMT and increase its background signal.
The PMT is also physically insulated from the combustion chamber by using poorly (thermally)
conducting metals to attach the PMT housing, filters, etc.
The physical arrangement of these components is as follows: flame (combustion) chamber with
exhaust, permanent thermal filter (two IR filters in some commercial designs), a removable
phosphorus or sulfur selective filter, and finally the PMT.

2.Photo ionization detector:


Schematic Diagram:

The selective determination of aromatic hydrocarbons or organo-heteroatom species is the job


of the photoionization detector (PID).

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This device uses ultraviolet light as a means of ionizing an analyte exiting from a GC column.
The ions produced by this process are collected by electrodes.
The current generated is therefore a measure of the analyte concentration.
The lamp energies range from 8.3 to 11.7ev and wavelength range from 150nm to 106nm
3. Electron capture detector.
Schematic Diagram:

The ECD uses a radioactive Beta emitter (electrons) to ionize some of the carrier gas and
produce a current between a biased pair of electrodes.
When organic molecules that contain electronegative functional groups, such as halogens,
phosphorous, and nitro groups pass by the detector, they capture some of the electrons
and reduce the current measured between the electrodes.
The ECD is as sensitive as the FID but has a limited dynamic range and finds its
greatest application in analysis of halogenated compounds.

UNIT III INDUSTRIAL GAS ANALYSER AND POLLUTION MONITORING


INSTRUMENTS

1. Draw and explain the measurement of dust and smoke in detail.(16) (Dec 2014, May 2013,
Nov/Dec 2016)
1. Electrostatic method of measurement

2. Optical type measurement


EI6501 Analytical Instruments Dept. of EIE and ICE 2016-2017

(1) Adapterthe smokemeter optical unit may be mounted on a fixed or movable frame. The
normal unrestricted shape of the exhaust plume shall not be modified by the adaptor, the meter,
or any ventilatory system used to remove the exhaust from the test site.
(2) Smokemeter (light extinction meter)continuous recording, full-flow light obscuration meter.
(i) It is positioned so that a built-in light beam traverses the exhaust smoke plume at right angles
to the axis of the exhaust stream.
(ii) The smokemeter light source shall be an incandescent lamp with a color temperature range
of 2800K to 3250K, or a light source with a spectral peak between 550 to 570 nanometers.
(iii) The light output is collimated to a beam with a maximum diameter of 1.125 inches and
an included angle of divergence within a 6 included angle.
(iv) The light detector shall be a photocell or photodiode. If the light source is an incandescent lamp,
the detector shall have a spectral response similar to the photopic curve of the human eye (a
maximum response in the range of 550 to 570 nanometers, to less than 4 percent of that maximum
response below 430 nanometers and above 680 nanometers).
(v) A collimating tube with apertures equal to the beam diameter is attached to the detector
to restrict the viewing angle of the detector to within a 16 included angle.
(vi) An amplified signal corresponding to the amount of light blocked is recorded continuously on
a remote recorder.
(vii) An air curtain across the light source and detector window assemblies may be used to
minimize deposition of smoke particles on those surfaces provided that it does not measurably
affect the opacity of the plume.
(viii) The smokemeter consists of two units; an optical unit and a remote control unit.
(ix) Light extinction meters employing substantially identical measurement principles and producing
substantially equivalent results, but which employ other electronic and optical techniques, may be
used only after having been approved in advance by the Administrator.
(3) Recordera continuous recorder, with variable chart speed over a minimal range of 0.5 to
8.0 inches per minute (or equivalent) and an automatic marker indicating 1-second intervals
continuously records the exhaust gas opacity, engine rpm and throttle position.
(i) The recorder is equipped to indicate only when the throttle is in the fully open or fully
closed position.
(ii) The recorder scale for opacity is linear and calibrated to read from 0 to 100 percent opacity
full scale.
(iii) The opacity trace has a resolution within one percent opacity.
(iv) The recorder scale for engine rpm is linear and has a resolution of 30 rpm.
(v) The throttle position trace clearly indicates when the throttle is in the fully open and fully
closed positions.
(vi) Any means other than a strip-chart recorder may be used provided it produces a permanent
visual data record of quality equal to or better than that described above (e.g., tabulated data,
traces, or plots).
(4) The recorder used with the smokemeter shall be capable of full-scale deflection in 0.5 second
or less. The smokemeter-recorder combination may be damped so that signals with a frequency
higher than 10 cycles per second are attenuated. A separate lowpass electronic filter with the
following performance characteristics may be installed between the smokemeter and the recorder
to achieve the high-frequency attenuation:
(i) Three decibel point10 cycles per
second. (ii) Insertion losszero 0.5 decibel.

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(iii) Selectivity12 decibels per octave above 10 cycles per second.


(iv) Attenuation27 decibels down at 40 cycles per second minimum.
(5) In lieu of the use of chart recorders, automatic data collection equipment may be used to record
all required data. Automatic data processing equipment may then be used to perform the data
analysis. The automatic data collection equipment must be capable of sampling at least two
records per second.
(c) Assembling equipment.
(1) The optical unit of the smoke meter shall be mounted radially to the exhaust pipe so that the
measurement will be made at right angles to the axis of the exhaust plume. For an end-of-line
smoke meter the distance from the optical centerline to the exhaust pipe outlet shall be 1 0.25 inch.
The full flow of the exhaust stream shall be centered between the source and the detector apertures
(or windows and lenses) and on the axis of the light beam.
(2) Power shall be supplied to the control unit of the smoke meter in time to allow at least
15 minutes for stabilization prior to testing.

2. Write short notes on Air pollution due to (Dec 2014)


a) Carbon monoxide (4)
b) Nitrogen oxide (4)
c) Hydrocarbons (4)
d) Sulphur dioxide (4)

1. Carbon monoxide:
Health Effects: Carbon monoxide enters the bloodstream through the lungs and reduces oxygen
delivery to the body's organs and tissues. The health threat from carbon monoxide at low levels is
most serious for those who suffer from cardiovascular disease, such as angina pectoris. At much
higher levels, carbon monoxide can be poisonous. Even healthy people may be affected. Visual
impairment, reduced work capacity, reduced manual dexterity, poor learning ability and difficulty in
performing complex tasks are all associated with exposure to carbon monoxide.
Sources: Motor vehicles produce about 60 percent of carbon monoxide nationwide; in cities, it may
be as high as 95 percent. Other sources include industrial processes, non-transportation fuel
combustion, and wildfires.
2. Nitrogen oxide:
Health Effects: Short-term exposure may cause increased respiratory illness in young children and
harm lung function in people with existing respiratory illnesses. Long-term exposure may lead to
increased susceptibility to respiratory infection and may cause alterations in the lung. (Nitrogen
oxides also can be transformed in the atmosphere to ozone of fine particulate soot - which are both
associated with serious adverse health effects.)
Environmental Effects: Nitrogen oxides help form acid rain. In addition, this pollutant can cause a
wide range of environmental damage, including visibility impairment and eutrophication - that is,
explosive algae growth which can deplete oxygen in water bodies such as the Chesapeake Bay.
Sources: Cars, trucks and electric power plants are dominant sources. Home heaters and gas stoves
can produce nitrogen dioxide inside homes.
3. Hydrocarbons:
In industrial areas and where there is heavy vehicular traffic, these contain carbon and
hydrocarbons from the incomplete combustion of fuels. The hydrocarbons include a variety of
polycyclic aromatic hydrocarbons (PAHs) that have been shown to cause mutations [
Inhaled particles smaller than 2.5 m ("PM2.5") are taken deep into the lungs and may be deposited
there. Chronic exposure to these small particles has been linked to
reduced lung development and function in adolescents;
an increased risk of asthma;
increases in the incidence of heart disease and lung cancer later in life;

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a significant increase in the frequency of germline mutations in the sperm of exposed mice.
4. Sulphur dioxide:
Health Effects: High concentrations of sulfur dioxide (SO2) can result in breathing problems with
asthmatic children and adults who are active outdoors. Short-term exposure has been linked to
wheezing, chest tightness and shortness of breath. Other effects associated with longer-term
exposure to sulfur dioxide, in conjunction with high levels of particulate soot, include respiratory
illness, alterations in the lungs' defenses and aggravation of existing cardiovascular disease.
Environmental Effects: Sulfur dioxide and nitrogen oxides are the major precursors of acid rain,
which has acidified soils, lakes and streams, accelerated corrosion of buildings and monuments, and
reduced visibility. Sulfur dioxide also is a major precursor of fine particulate soot, which poses a
significant health threat.
Sources: Combustion of fuel containing sulfur -- mostly coal and oil. Also produced during metal
smelting and other industrial processes.
3. Explain the working principle of Paramagnetic Oxygen Analyzer with a functional diagram.
Specify the need of oxygen measurement.(16) (May 2013,2014)

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4. With a Schematic Diagram explain the method of measuring Sulphur dioxide SO 2 estimation
using conductivity method(16) (May 2014 , May/ June 2016 )

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5. With a block diagram, explain the method of measuring carbon monoxide using Non-dispersive
Infrared Analyzer. (Nov 2013, May 2015, Nov/Dec 2016)
Colourless, odorless, poisonous gas that has affinity for haemoglobin which is 210 times that of
oxygen.
Hazardous pollutant, healthy individuals get affected.
It is a by-product of combustion processes in which incomplete oxidation of fossil fuels takes
place.
Associated with automotive exhaust and deep mining operations
Average concentration of this gas found to be below 200ppm.

6. Explain how the NO2 analyzer is used. (May 2013, May/ June 2016)
Nitrogen oxides formed when fuel is burned at very high temp (above 1200F) such as in
automobiles and power plants.
Nitric oxide and Nitrogen dioxide are main pollutants.
Nitrogen dioxide is a highly toxic, reddish brown gas that is formed through the oxidation of
nitric oxide emitted primarily form combustion of fuels.
It causes an odorous, brown haze that irritates the eyes
Emission of nitrogen oxides from the products of fuel combustion in furnaces and engine.
Its level ranges from 0.5 to 0.12 ppm on an annual average basis.

7. Explain how the H2S analyzer is used. (Nov/Dec 2016).

Hydrogen sulphide is highly toxic gas which comes out as an effluent from the industries. Lead
acetate tape staining method is used to find out the concentration of H2S in the flu gas.

Principle:When H2S reacts with lead acetate, it produces a brown patch of lead sulphide. This
patch when exposed to light absorbs radiation. The absorption of radiation is the measure of H2S
present in the flu gas.

Schematic of Lead acetate tape staining:

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Construction:
1. The equipment consists of a humidifier containing 5% acetic acid in water to humidify the H2S
2. A lead acetate tape is made to pass through a reaction window, where it is exposed to H2S
and light. This tape is driven by a motor
3. A measuring photocell and the sample photocell have been installed to measure the radiation
levels
4. A display device is deployed to give the readouts of the concentration of H2S

Working:

1. The H2S gas is humidified by acetic acid solution.


2. The humidified H2S is allowed pass through the lead acetate tape via a reaction window.
3. The H2S reacts with lead acetate tape and produces lead sulphide which is brown in color.
4. The brown patch will give the measure of H2S. To find out the concentration of H2S, a light
beam is passed over the brown patch and it is collected by a photocell.
5. The difference in intensities of radiation levels of reference beam and sample beam gives the
measure of H2S present in the solution.

8. Explain how the IR analyzer is used.


The Infrared radiation can be effectively used to analyze the presence of diatomic components such
as O2, N2 present in the flu gas. The vibration energy of these diatomic components matches with
the IR radiation wave length.
Principle: The diatomic molecules present in the sample absorb infrared radiation when exposed to
it. Hence there will be a reduction in the intensity of the IR radiation at the detector. The amount of
decrease in intensity is the measure of the amount of diatomic components present in the sample.
Construction:

The IR gas analysers consists of IR light source, a motor controlled chopper, a reference cell, a
sample cell and detector

The Motor controlled chopper is used to produce electric pulses rather than a continuous
beam of light

The reference cell consists of gases which do not absorb IR radiation

The sample cell has two openings. One for gas inlet and another is for gas outlet

The sample gas is made to pass through the sample continuously through these openings

An IR radiation detector is installed to capture the radiation passing out from the sample cell

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and the reference cell
Schematic of IR gas analyser:

Working:

a. The IR radiation is passed through the chopper and pulses of radiation are produced
b. This radiation is passed through the sample cell and the reference cell
c. The reference cell do not absorb IR radiation and hence the intensity of IR radiation will not
change
d. The diatomic molecules present in the sample absorb the IR radiation and hence the intensity of
the radiation will be reduced
e. The detector produces an appropriate electrical signal for the amount of radiation it
received. This electrical signal will be the difference in intensity levels coming out from reference
cell and the sample cell. The electrical signal will be the measure of diatomic components present in
the sample
9. With a suitable diagram explain the construction and working principle of thermal
conductivity analyzer. (Nov 2013) (May/ June 2016)
Thermal conductivity analyzer makes use of thermal conductivity principle to measure amount of
gas to be measured in the flu gas.

Principle: When a cooled gas is made to pass through the heated metal connected to a power
supply, the gas will absorb the heat. This will change the current flow through the metal. It will give
the measure of the gas.

Schematic of Thermal Conductivity Analyzers

Construction:

1. The thermal conductivity sensors consist of a wheat stone bridge


2. Two arms of the bridge contain heated platinum wires in opposite direction over which the
sample and the reference gases are passed
3. An external power supply is used to excite the bridge
4. The output from the bridge is connected to amplifier and to signal conditioning circuit

Working:

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1. The bridge has to be balanced before measuring the gas of our interest
2. Initially the reference gas is allowed to pass over the arms and the current flow through the
bridge is noted
3. If the galvanometer shows zero reading then the bridge is balanced. If the bridge is not balanced
external voltage is applied and the bridge is brought back to balanced condition
4. After attaining balance condition in the bridge, the sample gas is passed over one of the arms of
the bridge containing heated platinum wire.
5. The sample gas absorbs the heat from platinum wire which in turn causes change in current flow
through the wire and hence the bridge balance is lost
This results in output Voltage and it is the measure of the sample gas concentration.
10. What are the sources of air pollution and explain in detail?
Answer in Question No.2
11. What are the types of gas analyzers? Explain anyone with example.
A. Paramagnetic oxygen Analyzer
B. Zirconia Oxygen Analyszer
C. Infrared gas analyzer
D. Tunable diode laser
E. Thermal conductivity Analyzer.

PARAMAGNETIC OXYGEN ANALYSER

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THERMAL CONDUCTIVITY ANALYZERS

Thermal conductivity analyzer makes use of thermal conductivity principle to measure amount of
gas to be measured in the flu gas.

Principle: When a cooled gas is made to pass through the heated metal connected to a power
supply, the gas will absorb the heat. This will change the current flow through the metal. It will give
the measure of the gas.

Schematic of Thermal Conductivity Analyzers

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Construction:

5. The thermal conductivity sensors consist of a wheat stone bridge


6. Two arms of the bridge contain heated platinum wires in opposite direction over which the
sample and the reference gases are passed
7. An external power supply is used to excite the bridge
8. The output from the bridge is connected to amplifier and to signal conditioning circuit

Working:

6. The bridge has to be balanced before measuring the gas of our interest
7. Initially the reference gas is allowed to pass over the arms and the current flow through the
bridge is noted
8. If the galvanometer shows zero reading then the bridge is balanced. If the bridge is not balanced
external voltage is applied and the bridge is brought back to balanced condition
9. After attaining balance condition in the bridge, the sample gas is passed over one of the arms of
the bridge containing heated platinum wire.
10. The sample gas absorbs the heat from platinum wire which in turn causes change in current flow
through the wire and hence the bridge balance is lost
11. This results in output Voltage and it is the measure of the sample gas concentration.
12. With the instrumentation setups explain any one method to estimate sulphur-di-oxide present in air.
(May/ June 2016)
A. The Colorimetric ripper method.
B. The Potentiometric method.
C. The aeration oxidation method.
D. Photometric Method
1. Absorbance
2. Reflectance
13. Explain the photometric method of measuring dust and smoke measurement in thermal power plant.
Answer in Question No:1
14. Why do we need temperature compensation in conductivity measurement? Describe a method of
temperature compensation
Conductivity is a function of temperature of the solution. Increase in temperature causes increase in
mobility of electrons and thus increases the conductivity. But the conductivity of a solution is
defined for a particular temperature. Since the measurement of conductivity may not be done under
the specified temperature compensation circuit must be incorporated to compensate the impact of
temperature variations.

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15. Draw the schematic diagram of hotwire thermal conductivity analyzer and explain its operation (May 2015)
Thermal conductivity analyzer makes use of thermal conductivity principle to measure amount of
gas to be measured in the flu gas.

Principle: When a cooled gas is made to pass through the heated metal connected to a power
supply, the gas will absorb the heat. This will change the current flow through the metal. It will give
the measure of the gas.

Schematic of Thermal Conductivity Analyzers

Construction:

1. The thermal conductivity sensors consist of a wheat stone bridge


2. Two arms of the bridge contain heated platinum wires in opposite direction over which the
sample and the reference gases are passed
3. An external power supply is used to excite the bridge
4. The output from the bridge is connected to amplifier and to signal conditioning circuit

Working:

1. The bridge has to be balanced before measuring the gas of our interest
2. Initially the reference gas is allowed to pass over the arms and the current flow through
the bridge is noted
3. If the galvanometer shows zero reading then the bridge is balanced. If the bridge is not
balanced external voltage is applied and the bridge is brought back to balanced condition
4. After attaining balance condition in the bridge, the sample gas is passed over one of the arms
of the bridge containing heated platinum wire.
5. The sample gas absorbs the heat from platinum wire which in turn causes change in current flow
through the wire and hence the bridge balance is lost
This results in output Voltage and it is the measure of the sample gas concentration.

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Unit IV pH METER AND DISSOLVED COMPONENT ANALYZER

1. Explain in detail about construction and working principle of ammonia electrodes.(16) (Dec
2014)

PRINCIPLE:
An ion-selective electrode (ISE), also known as a specific ion electrode (SIE), is
a transducer (or sensor) that converts the activity of a specific ion dissolved in a solution into an
electrical potential, which can be measured by a voltmeter or pH meter. The voltage is
theoretically dependent on the logarithm of the ionic activity, according to the Nernst equation.
The sensing part of the electrode is usually made as an ion-specific membrane, along with
a reference electrode. Ion-selective electrodes are used in biochemical and biophysical research,
where measurements of ionic concentration in an aqueous solution are required, usually on a real
time basis.
WORKING:
Just like most common pH electrodes, ISEs work on principle of galvanic cell. By
measuring electric potential generated across a membrane by selected ions, and comparing it to
a reference electrode, a net charge is determined. Strength of this charge is directly proportional to
concentration of selected ion.
TYPES:
The 4 main types of ISEs are:
1. Glass Electrodes
2. Solid state Electrodes
3. Liquid-Liquid Membrane Electrodes
4. Gas Sensing Electrodes

GLASS ELECTRODES:
First of this kind was developed for H+ ions. Glasses having less than 1% Al2O3 are sensitive
only to H+ ions. Those glasses of composition Na2O-11% Al2O3-18% SiO2-71% is highly selective
towards sodium. Glass electrodes have been made selective to Na, K, NH3, Ag.
SOLID STATE ELECTRODES:

They use single crystals of inorganic material doped with a rare earth. They are
particularly useful for halide ion analysis.
LIQUID-LIQUID MEMBRANE ELECTRODES:

They are essentially liquid ion exchangers separated from the liquid sample by means of
a permeable membrane which allows liquids to come in contact with each other but prevents

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their mixing. Based on principle, ISEs sensitive to Ca and Mg have been developed.
GAS SENSING ELECTRODES:
They respond to partial pressure of the gases in sample. Gas is transferred across a gas
permeable membrane, until the partial pressure in thin film of filling solution between glass and
membrane and probe membrane equals in that sample. Resultant pH change is measured by a
combination pH electrode, a potential is developed related to the partial pressure and hence tha
ammonia or SO2 concn.
Problems:
The response of the electrode and galvanometer is temperature sensitive, and also 'drifts'
over time, requiring recalibration frequently during a series of measurements, ideally at least one
calibration sample before and after each test sample. On the other hand, after immersion in the
solution there is a 'settling time' which can be five minutes or even longer, before the electrode
and galvanometer equilibrate to a new reading; so that timing of the reading is critical in order to
find the most accurate 'window' after the response has settled, but before it has drifted
appreciably.
Advantages
1. Simple
2. Inexpensive
3. Robust
4. Durable
5. Easy usage
6. Accuracy and precision up to 2 or 3%
7. Measure both positive and negative ions
8. Unaffected by turbidity or sample

color Wide temperature range

2. Write short notes on: (Dec 2014, May 2015, Dec 2015)
Sodium Analyzer(8) (May 2013, May/ June 2016)

Sodium analyzers are mainly used in thermal power plants for the determination of sodium
ion concentration in the boiler water. The analysis of sodium is important, because the excess of

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sodium will corrode the material in which it is passing. The important salts of sodium that will
cause the corrosion are sodium hydroxide, sodium chloride and sodium sulphate.

This analysis is also helpful for the detection of leakage in the condenser. The leakage can be
detected by measuring the concentration of sodium in steam and in the condensate. If the two
measurements give the same value of sodium concentration then that indicates the leakage in the
condenser.

Preparation of sample is important in the analyzer. It has to be prepared well for the accurate
measurement of ion under study. The figure shown in 4.16 is the sodium analyzer. It consists of
overhead tank for storing the sample, standard solutions for calibrating the instrument, 3 way
solenoid valves for allowing either sample or the standard solution. Ammonia buffer for pH
adjustment, reference and sodium ion specific electrode for concentration measurement, amplifier
and indicator.

Normally the electrodes measure the activity of the ion. (i.e.) the potential developed by the
electrode is actually proportional to the logarithm of the ionic activity. The activity is a measure of
the free ions in the solutions. But our interest of study is to measure the concentration of the
solution [Both the free ions and the bound ions give the concentration].

So to measure the concentration the bound ions must be freed by adding reagents with it. The
addition of reagents not only liberates the bound (Na) ions but also prevents the unwanted ions
from entering ion measurement. The sodium selective electrode provides good response in the
measurement of sodium ion in buffered solutions having the pH value of above 1o. This pH value
could be achieved by adding ammonia buffer solutions with the sample.

The sample which is to be analyzed is maintained in the constant head tank. The sample is then
allowed to reach the flow cell where the reference and ion selective electrodes are placed. The
sample flow can be controlled by the three way solenoid valve.

The concentration of the sodium ion can be measured with the help of electrodes. The output of
the electrode is in term of potential which is proportional to the logarithm of the sodium ion
concentration in the solution .The potential value is amplified it can either be recorded or
indicated.

The efficiency of the system can be improved in two ways. They are
1.
Standardization
2.
Cleaning process

Standardization is the process of calibrating the instrument to read the correct value. This process
starts by activating a three way solenoid valve to stop the sample flow and to allow the standard
sodium ion solution into the flow cell. When the electrodes have stabilized in the new solution, the
amplifier output is compared with the standard value. If there is any deviation between the
measured and standard value, the output is adjusted to read the same standard value. After
standardization, the solenoid valve is adjusted to allow the sample solution for the measurement.

Cleaning the electrode is very important, because after the usage of the electrode for a particular
time period, the electrode surface gets the deposition of the unwanted materials in the solution and
it affects the sensing capability of the electrode. So for the accurate measurement of the sodium
ion, it is necessary to deposits off the electrode. Spray nozzle,

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Silicon Analyzer(8)

SILICA ANALYZER:

PURPOSE:

1. In thermal power plants, silica measurement boiler water is very important.

2. The need for finding silica content is that the silica present in water and steam has the
tendency to form deposition in the pipes and on turbine blades.

3. This kind of deposition affects the efficiency of the equipment, in turn affects the entire
power plant efficiency.

4. So continuous monitoring of silica contents is necessary to maintain the systems


efficiency.

PRINCIPLE:

1. The instrument used to measure the silica content is the silicometer.


2. It uses the colorimetric principle for the measurement.

3. The sample is prepared for analyzing by molybdate, sodium citrate, amino acids to the
sample.

4. These added chemicals react with the silica to give a reaction product of a specific colour.
The light absorbed of this product is measured with a photometer and is proportional to the
concentration of silica.

5. The figure shows a silica analyzer woks on the colorimetric principle which is based on
molybdenum blue method.

CONSTRUCTION AND WORKING:

The sample is prepared for analyzing by adding ammonium molybdate solution, sulpuric acid and
a reducing solution with the sample. These are mixed well in the mixing vessel with the help of
stirrer and the flow rate of each is controlled by the valve which is placed on each line. The inlet
of mixing vessel and the measuring cuvette is maintained at the top to prevent the contamination
by any solution from the previous analysis to avoid the interference with next analysis.

The analyzer is having two cuvettes, one lamp source and two photometers. The two cuvettes are
namely reference cuvette and measuring cuvette. The reference cuvette is used for holding the
reference solution having the known concentration of silica and the measuring cuvette is used for
holding the sample whose silica concentration is to be measured. The lamp source is placed in
between the measuring and reference cuvettes, the two photometers are placed on either side of
the two cuvettes.

After preparing the sample, the measuring cuvette is filled with the sample and the reference
cuvette is filled with the solution having known concentration of sodium ions. The light sources
are allowed to pass through both the measuring and reference cuvettes. The transmitted light rays
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are detected by the photo voltaic cells. The output of the two photo voltaic cells drives the
differential amplifier and the amplified signal is read through the read out devices. In this, if both
the reference solution and the sample have the same concentration of silica the output of the
differential amplifier is zero. The relationship between the light absorption, transmittance, photo
detector output, differential amplifier and concentration are given below.

3. What is a reference Electrodes? What are the various types of Reference Electrodes?
Compare the properties of different types of Reference Electrodes?(16) (May 2014)

CALOMEL ELECTRODE OR REFERENCE ELECTRODE

The purpose of the reference electrode is to provide a stable, reproducible voltage to which the
working electrode potential may be referenced. A reference electrode may be considered a small
battery whose voltage is determined by the chemistry taking place between a solid conductor and
the electrolytic solution around it. If a small current is passed through it, the potential change is
negligible.

The common reference electrodes are:


1. Mercury/mercurous electrode
2. Silver/silver chloride electrode

The reference electrode against which the potential of the glass electrode is measured is the
calomel electrode. It consists of, a metallic internal element, typically of mercury-mercurous
chloride or silver/ silver chloride, immersed in an electrolyte, which is usually a saturated solution
of potassium chloride.

The electrolytic solution forms a conductive salt bridge between the metallic element and the
sample solution. In which the measuring and reference electrodes are emplaced. For a stable
electrical connection between the internal metallic element and the sample solution, a small but
constant flow of electrolyte solution is maintained through a liquid junction in the tip of the outer
body of the reference electrode. Depending upon the nature of the application, this junction may
be formed in several ways. For example, the tip of the electrode could be formed by an embedded
linen or asbestos fibre, or by a permeable composition of pressed-sintered carborundum and glass
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pellets.

1/2Hg2Cl2+e Hg+Cl-

Since at a given temperature the activity of the mercurous chloride is constant and that of the
mercury is unity by definition, it is the chloride ion activity which is the potential determining.
When this is fixed, the electrode has a fixed potential at a fixed temperature. Although calomel
reference electrode is the most commonly used electrode, it is sometimes necessary to use other
types of reference electrodes in special circumstances. For ex.,

1. The Ag\AgCl electrode is recommended as a reference electrode at high temperature

2. The mercury/mercurous sulphate electrode with a salt bridge solution of potassium sulphate is
used in test solution that must not contain chloride ions.

3. The calomel electrode with open liquid junction facilitates measurements in contaminated
samples and samples containing solid particles.

4. The calomel electrode with a salt bridge solution of saturated lithium chloride is used for the
measurements in non-aqueous solutions.LiCl is more suitable in organic solvents than is KCl.

5. A calomel electrode with duplicate salt bridge may be used if test solution must not contain
chloride ions.

The reference electrode should be stored in a small beaker containing the salt bridge solution for
short term storage. For a long term storage, the electrode should be rinsed, dried and stored with
the end cap on and the rubber band covering the filling hole in place. The solution in the salt
bridge should always be on a higher level than the solution to be measured, as infiltration of the
sample solution may occur in the salt bridge.Annular ceramic junctions and porous pin junctions
can occasionally become blocked due to crystallization of the salt bridge filling solution.Soaking
in the salt bridge solution usually remedies the situation,but,on those instance that it does not
,raising the temperature to the maximum allowable for the reference system will often
help.Trapped bubbles can also cause malfunctions.these bubbles can be removed by gently
tapping the electrode or shaking it downward

SILVER/SILVER CHLORIDE REFERENCE ELCTRODE

The silver/silver chloride (Ag/AgCl) electrode features glass-body construction with a porous
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ceramic frit at the solution interface. The filling solution is 3M NaCl gel that has been saturated
with AgCl, the gel is semi solid.

A porous reference junction separates the filling solution in the electrode from solution whose pH
is to be measured. The filling solutions constant chloride ion concentration generates potential at
a pure silver wire with silver chloride on it. The silver wire passes the signal from the solution
being measured to the electrodes cable or connector. This configuration of the electrode is called
SINGLE JUNCTION ELECTRODE.

If samples contain proteins,sulfides,heavy metals or any other material which interacts with silver
ions,they may react with gel,causing a reduction in the reference output.this reaction can lead to
erroneous reference signals or to precipitation at the reference junction leading to a short service
life. In a double junction reference electrode, the inner chamber contains the usual high salt
concentration solutions so that stable outputs are generated. The outer chamber, which contacts the
sample through the porous reference junction,is filled with0.1M KCl.This lower ionic strength
material more closely matches that of the sample and further reduces spurious potentials.

The Ag/AgCl reference electrodes are easily spoiled by drying.It is,therefore,advisable to keep the
tips wetted at all times and store in 3M NaCl when not in use.This helps to extend its life time.
They usually last 3-6 months.

HYDROGEN ELECTRODE

It comprises a platinum plate covered with platinum black kept in the hydrogen element, gaseous
hydrogen at atmospheric pressure acting directly on this plate.

It consists of a hollow tube which is opened at the bottom and is immersed in the solution whose
pH value is to be measured. The platinum plate is inserted into this hollow tube and is joined to
the platinum wire which is welded to the glass tube,this platinum wire is joined to a copper
lead.The hydrogen is fed to the other tube from above and passes out through the lower aperture
placed at level,half way up the platinum plate.the hydrogen electrode is joined to the comparison
half cell by an electrolytic solution.the cell is filled with a n inert salt and closed bya semi-
permaeble stopper.this means of connecting the elements lowers the diffusion potential ,which
arises at the boundary of the two solutions and introduces an error into the result of the
measurement.

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The standard hydrogen cell with the normal concentration of hydrogen ions can be employed as
the comparison cell.The potential of the standard hydrogen electrode is tentatively taken as zero,in
relation
to which the potentials of the other electrodes are determined. Thus E0=0,the e.m.f of a measuring
cell with hydrogen electrodes will be equal to E=0.0001982(273+t).(-logaH+)
Where aH+ is the active concentration of hydrogen ion of the solution being measured.
pH=E/(0.0001982(273+t)
this method is most likely used in labs for pH measurements from 1 to 14.its use is limited
application because of some of the operation defects which are:
1.necessity of saturating with hydrogen.
2. poisoning of electrode in consequence of the absorption by the platinum black of some
component parts of the solution.

4. State the principles of measuring Dissolved Oxygen in Boiler Feed Water. Explain any one

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method to measure the dissolved oxygen.(16) (May 2014)

Refer : Q.No:8
5. Describe how a conductivity cell is used. Give the applications of conductivity measurement
of a liquid. Explain how the oxygen analyzer is used. (May/ June 2016)

Thermal conductivity analyzers


a) Thermal conductivity of a gas is defined It is defined as the quantity of heat
(calories) transferred in unit time (secs) in a gas between two surfaces 1 sq.cm in
area and 1 cm apart with a temperature difference between the surface 1 deg C.
b) A gas analysis based on the thermal conductivity includes a measuring component
whose thermal conductivity differs sufficiently from the thermal conductivity to the
carrier gas.
c) In a typical hot wire cell thermal conductivity analyser, four platinum filaments are
employed as heat sensing elements.

Two filaments connected in opposite arms of the Wheatstone bridge act as reference arms, other
two filaments are connected in the gas stream.

stream passed through the measuring pair of filaments, the wire cools and resistance change.
ld be the resistance.
Analysis based on ionization of gases.

gas in a mixture.
is ionized,
selectively filtered and detected with a photocell.

6. Describe the construction of pH electrode. Draw the electronic circuit diagram for

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measuring pH of a liquid and explain its working. (Nov/Dec 2016)


pH is a unit of measure which describes the degree of acidity or alkalinity (basic) of a solution.

It is measured on a scale of 0 to 14.

The formal definition of pH is the negative logarithm of the hydrogen ion activity.

pH = -log[H+]

The pH value of a substance is directly related to the ratio of the hydrogen ion and hydroxyl ion
concentrations.

If the H+ concentration is higher than OH- the material is acidic.

If the OH- concentration is higher than H+ the material is basic.

7 is neutral, < is acidic, >7 is basic

The pH scale corresponds to the concentration of hydrogen ions.

For example pure water H+ ion concentration is 1 x 10-7 M, therefore the pH would then be 7.

The addition of acid to water increases the concentration of hydrogen ions and reduces the
concentration of hydroxyl ions

The addition of a
base would increase the concentration of hydroxyl ions and decrease the concentration
of hydrogen ions

An acid canbe defined as a proton donor, a chemical that increases the concentration of hydrogen ions
in solution.

A base can be defined as a proton acceptor, a chemical that reduces the concentration of hydrogen ions
in solution.

7. Discuss how pH values are measured. Explain the role of calomel electrodes in this
measurement.

A pH measurement system consists of three parts: a pH measuring electrode, a reference


electrode, and a high input meter.The pH measuring electrode is a hydrogen ion sensitive glass
bulb.The reference electrode output does not vary with the activity of the hydrogen ion.
A sample is placed in a cup and the glass probe at the end of the retractable arm is placed in it.

The probe is connected to the main box.

There are two electrodes inside the probe that measure voltage.
One is contained in liquid with fixed pH.
The other measures the acidity of the sample through the amount of H+ ions.
A voltmeter in the probe measures the difference between the voltages of the two electrodes.
The meter then translates the voltage difference into pH and displays it on the screen.
Before taking a pH measurement the meter must be calibrated using a solution of known pH.

Temperature compensation is contained within the instrument because pH electrodes are


temperature sensitive.Temperature compensation only corrects for the change in the output of
the electrode, not for the change in the actual solution.
EI6501 Analytical Instruments Dept. of EIE and ICE 2016-2017

8. Briefly explain about dissolved oxygen analyzer.( Nov/Dec 2016)

The paramagnetic oxygen analyzer based on the phenomenon of magnetic wind, which
rises due to the motion of paramagnetic oxygen molecules in to the non-uniform magnetic field.
The flowing gas is made to pass over a heated filament connected as one arm of a Wheatstone
bridge circuit. The gas cools the filament and the unbalances the bridge circuit. The output
voltage from the bridge can be fed to a recorder, to show a continuous indication of oxygen
concentration in the sample.
The Hartman and Braun oxygen analyzer method use of the dependency of paramagnetic
susceptibility on the temperature according to the curie law. The analyzer comprises a ring
chamber which consists of a metallic hollow ring arranged in vertical plane, with a gas inlet
below and a gas out let above. The gas is conducted in the upward direction. The two halves of
the ring chamber are connected by a glass tube. The transverse tube has taped wire winding on
its outside. The winding arms a part of a Wheatstone bridge circuit, whose one-half is placed in
the field of the permanent magnet.
In case the sample gas does not contain any oxygen, there is no flow in the horizontally placed
transverse tube. If, however the sample contains oxygen, it is attracted into the magnetic field.
This gas flow is heated and due to increase in the temperature, the susceptibility of the
paramagnetic oxygen molecules decreases. In this way, the heated gas is pushed away by the
cold gas coming from the left hand side. A gas flow thus arises inside the transverse tube. This
flow is called magnetic wind, whose velocity depends upon the oxygen concentration in the
sample gas. The magnetic wind cools down the left side of the heated winding more than the
right side.
Therefore, a change in the bridge balance takes place, which depends upon the temperature
gradient. The voltage difference of the bridge is proportional to the oxygen concentration in the
measuring gas.
The instrument suffers from the following sources of the error:
1) The filament is affected by change in the thermal conductivity of the carrier gas. Thus,
the calibration is correct for only one gas mixture, which must be said for each analyser.

2) Hydrocarbons and other combustible gases in the sample stream react in the heated
filaments causing changes in temperature and therefore their resistance causes large
errors.

3) The cross tube must be horizontal to avoid an error due to gravitational chimney flow
effects

The electrochemical methods:


The methods that are most commonly used to measure dissolved oxygen (DO) can be sorted into
three major groups: colorimetric, titrimetric, and polarographic. Because oxygen can easily
diffuse into water sample during handling and alter the concentration of dissolved oxygen,
samples have to be collected and processed without contact with air.
Colorimetric: The sample fluid is sucked into an ampoule under vacuum (an ampoule is a sealed
glass tube with an easy-to-break tip. Use of the ampoule involves breaking of the tip). Chemical
reagents that are present in the ampoule in excess interact with oxygen to form a colored product
(that absorbs light at a visible wavelength). The intensity of the color, which is proportional to the
oxygen concentration in that sample, is compared to a series of tubes with color intensities that
reflect known concentrations of dissolved oxygen and are expressed in units of mg/l. Colorimetric
kits provide for rough and rapid measurement: they are often used for screening for low-oxygen
conditions and can be easily used by non-professional operators.
Titrimetric: A water sample is trapped in a dissolved oxygen bottle, which is a bottle with

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specially-designed cap (or specially-designed mouth and glass stopper, often sold as BOD
bottle), that allow for enclosure of liquids without contact with air. Chemical reagents added in
excess interact with oxygen to form a product, and another chemical (the titrant) is used
quantitatively to neutralize that product. The amount of titrant needed is proportional to oxygen
concentration and is translated to mg/l. The best known titrimetric method for dissolved oxygen is
the Winkler method; this method and its modifications are widely applied, both in the field (using
a syringe to dispense the titrant) and in the lab (using high-precision burettes). Field kits are useful
for routine monitoring of D.O. in creeks. Samples can be collected and fixed in the field, and
titrated later at home or in the lab. Laboratory applications include D.O. electrode calibration.

Polarographic: Electrodes measure the flux of oxygen across a membrane. Oxygen is consumed
in the process, and the traditional electrodes require flushing of measured liquid at the membrane
surface to constantly replace the oxygen consumed. A new technology provides Rapid-Pulse
probes with intermittent operation period that are so short they consume only a negligible amount
of oxygen; these probes are used in automatic/remote sensors with data loggers. The output of DO
electrodes is expressed in mg/l. Electrodes are useful for measurements of D.O. along gradients or
transects where many samples are needed in a short time, measurements of kinetics of change in
D.O. concentrations, continuous monitoring of D.O. with data-logging, and micro-scale D.O.
gradients on sediment surface (microelectrodes).

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9. Discuss in detail the principle, characteristics of electrodes used in pH meters.

The convention is to designate the cell so that the standard hydrogen electrode
is written first. H2(g) 2H+(aq) + 2e-
The electrode is used as a reference
(of zero) for the values of other standard electrode potentials.
Electrode life 7-20 days
Pressure 1 atm
Placed in a solution of 1M Hcl
Advantages
1. It is standard reference electrode with which the potentials of other electrodes are
calculated.
2. The results are highly accurate.
3. It gives no salt error.
4. Error due to electrical leakage is negligible.
Dis Advantages
1. Platinum is only poised by the absorption of impurities from the solution and the gas.
2. The presence of oxidising agents,unsaturated organic compounds, alkaloids, ions of noble
metals etc in the solution displaces the equilibrium and alters the potential.
3. The hydrogen gas at 1 atm pressure is difficult to set up and transport.
10. Discuss how pH of a solution is measured using glass electrode and reference electrode with
necessary diagrams. Refer : Q.No:9
11. Explain about sodium analyzer in detail. (May 2013)Refer : Q.No:2
12. Explain about silica analyzerRefer : Q.No:2
13. Explain different types of ion selective electrodes. (Nov 2013) Refer : Q.No:1
14. List the types of electrodes used for pH measurement. Explain the construction details of
Digital pH meter. Also specify the need of reference electrode for pH measurement.Refer :
Q.No:9
15. Describe a method of measuring dissolved oxygen content in the boiler feed water. Why
dissolved oxygen content is monitored in feed water? Also specify the cause of dissolved
oxygen content in feed water. (May 2013)
Refer : Q.No:4
16. Explain the working principles of biosensors with a conceptual diagram. Also specify the
various parameters to be measured using Biosensor. (Nov 2013, May/ June 2016)

BIO SENSOR

A biosensor is a device that uses biological materials to monitor the presence of various chemicals
in a substance or it can also be defined as it is a device for the detection of an analyte that
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combines a biological component with a physiochemical detector component.

Features of bio sensor


A good bio sensor must possess at least some of the following features. They are

1. The biocatalyst which is used in biosensor must be highly specific for the purpose of the
analyses and should be stable under normal storage conditions.

2. The reaction should be independent of the physical parameters such as pH, temperature etc.

3. The response should be accurate, precise, reproducible and linear over the useful analytical
range.

4. It should be free from electrical noise.

5. In clinical situations, for invasive monitoring, the bio sensor used must be tiny and bio
compatible, having no toxic or antigenic effects.

6. The complete bio sensor should be cheap, small, portable and capable of being used by Semi-
skilled operators.

7. If the reaction involves cofactors or coenzymes, these should also be co-immobilized with the
enzyme.

Block Diagram of Bio Sensor

Biosensors are modification of the ion selective electrodes, in which an enzyme is immobilibsed
on the surface of the glass electrode. So the biosensor is a combination of biological and physio -
chemical components. The biological component can consist of enzymes, antibodies, whole cells
or tissue slices and is used to recognize a specific analyte. The physio - chemical component often
referred to as the transducer which converts this interaction into a signal. This can be amplified
processed and read out by a device. A best example of the biosensor is the urea or the ammonia
sensing electrode.

The glucose sensor is also a good example for biosensor. It is used for measuring the blood
glucose in diabetic patients. It combines glucose oxidase with an electrode to measure the oxygen
generated by the reaction. The figure shown is the glucose sensor with an oxygen electrode. It
works on the principle that oxygen is consumed in the presence of glucose, providing a change in
the signal from the conventional oxygen electrode.

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In the construction of glucose sensor the immobilized enzyme is held between the inner and outer
membrane electrode. The gel used in polyacrylamide. This sensor just responds for the presence of
glucose by changing the value of the oxygen. The measure of change in oxygen value is a measure
of the glucose value. The output of the sensor can be converted into electrical signal by a
transducer, amplified, processed and displayed. These sensors are primarily used in food and
beverage industries

17. Explain the operating principle of kathrometer in measuring dissolved oxygen (May 2015,
Nov/Dec 2016)
The dissolved oxygen in feed water is a corrosion accelerator. The unit used to mention
dissolved oxygen is ppm. Analysis to quantify oxygen can be done either by laboratory
method or by on-line analysis. Chemical titration method is normally used to find out the
quantity of dissolved oxygen from the sample collected and immediately presented for the test.
The validity of measurement will be under suspicion as the delay causes absorption of oxygen
from atmosphere to sample. Special portable kits are used to carry out the testing on the spot.
Its always better to use analyzers that can test online. Kathrometer is of the types.when the
sample is cooled and scrubbed by hydrogen,the mixed oxygen in the water will be displaced to
form a mixture of hydrogen and oxygen. Wheatstones bridge circuit is used to measure the
amount of dissolved oxygen. The heated wires of bridge is exposed to hydrogen in two
opposite arms and to oxygen-hydrogen mixture in the opposite two arms. The unequal cooling
of opposite arms filaments makes the bridge unbalanced.
The unbalanced voltage can be calibrated in terms of dissolved oxygen. The kathrometer has
four major parts to perform the above function, the cooler, scrubber, hydrogen producer and
wheatstones bridge.

18. Explain the operating principle of solid state sensors and liquid matrix electrodes with neat
diagrams ( Dec 2015)
a)Solid state sensors
In these devices the ion sensitive material is bonded with FET. The requires the material and its
method of fabrication to be compatible with substrate(high purity silicon). This very significant
requirement puts a severe limitation on the use of some of the best-characterised membrane
materials, including ion sensitive gases. Martin and Sinclair report pH sensitive electrode by thick
film screening technique. This electrode retain the property of ion sensitive FET transducer but
eliminates the restrictions on membrane selection and fabrication. Here ion sensitive structure is
physically separated from the FET. In this way ion sensitive membrane can be fabricated on a
compatible substrate and the FET can then be attached appropriately and placed in close proximity
of ion-sensitive membrane. A hybrid electrode structure permits the incorporation of a source
follower FET amplifier, directly adjacent to the pH membrane, significantly reducing response
time and noise pick up
The below figure presents the cross section of an ISFET, which is essentially a conventional
insulated gate field effect transistor, that has its metallic gate contact replaced by a chemically
sensitive coating and a reference electrode. In solution, the gate region can be coated with an ion-
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sensitive membrane. Interaction of ions in solution with the membrane results in a change of
interfacial potential and corresponding alteration of drain current. By this technique, numerous
cations and anions have been sensed. The ISFET has advantages in its small size and low output
impedance which makes it ideal for invivo monitoring or analysis of small sample volumes.
However, problems like ion-selective coating adhesion and device encapsulation have prevented
large scale use of ISFETs. Other examples are CHEMFETs, ion-controlled diodes, schottky
diodes, thin film tin oxide gas sensors and chemiresistors.

UNIT V ELECTRO MAGNETIC RESONANCE AND MICROSCOPIC TECHNIQUES

1. Explain Scanning Electron microscope in detail and also give its applications.(16) (Dec 2014)
SCANNING ELECTRON MICROSCOPY
(SEM) 1. Introduction
The SEM instrument is made up of two main components, the electronic console and the electron
column. The electronic console provides control knobs and switches that allow for instrument
adjustments such as filament current, accelerating voltage, focus, magnification, brightness and
contrast. The FEI Quanta 200 is a state of the art electron microscope that uses a computer system in
conjunction with the electronic console making it unnecessary to have bulky console that houses
control knobs, CRTs, and an image capture device. All of the primary controls are accessed through
the computer system using the mouse and keyboard. The user need only be familiar with the GUI or
software that controls the instrument rather than control knobs and switches typically found on older
style scanning electron microscopes. The image that is produced by the SEM is usually viewed on
CRTs located on the electronic console but, instead with FEI the image can be seen on the computer
monitor. Images that are captured can be saved in digital format or printed directly.

The electron column is where the electron beam is generated under vacuum, focused to a small
diameter, and scanned across the surface of a specimen by electromagnetic deflection coils. The lower
portion of the column is called the specimen chamber. The secondary electron detector is located
above the sample stage inside the specimen chamber. Specimens are mounted and secured onto the
stage which is controlled by a goniometer. The manual stage controls are found on the front side of the
specimen chamber and allow for x-y-z movement, 360 rotation and 90 tilt however only the tilt cannot
be controlled through the computer system thus there is no need to use all of the manual controls
manipulate the orientation of the sample inside the sample chamber.
Below is a diagram of the electron column and a description of each of the components of the electron
column.

MEDICINE

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I. It is used to study about the viruses which acts as a disease causing agents. A knowledge of their
structure helps to find the methods of their destruction.
II. It is used to study about the bacteriaes in a detail
manner. a. INDUSTRY
III. It is used to find the chemical compositions of alloys, metals, semiconductors etc.
IV. The surface structure or topography can be studied easily with SEM.

2. Explain Transmission Electron Microscope in detail and also give its applications.(16) (Dec 2014)
Transmission Electron Microscopy
A transmission electron microscope is constituted of: (1) two or three condenser lenses to focus the
electron beam on the sample, (2) an objective lens to form the diffraction in the back focal plane and
the image of the sample in the image plane, (3) some intermediate lenses to magnify the image or the
diffraction pattern on the screen. If the sample is thin (< 200 nm) and constituted of light chemical
elements, the image presents a very low contrast when it is focused. To obtain an amplitude contrasted
image, an objective diaphragm is inserted in the back focal plane to select the transmitted beam (and
possibly few diffracted beam): the

crystalline parts in Bragg orientation appear dark and the amorphous or not Bragg oriented parts
appear bright. This imaging mode is called bright field mode BF (Fig. 3.7). If the diffraction is
constituted by many diffracting phases, each of them can be differentiated by selecting one of its
diffracted beams with the objective diaphragm. To do that, the incident beam must be tilted so that the
diffracted beam is put on the objective lens axis to avoid offaxis aberrations (Fig. 3.8). This mode is
called dark field mode DF. The BF and DF modes are used for imaging materials to nanometer scale.

Scanning Transmission Electron Microscopy STEM


The basic principle of image formation in the STEM is fundamentally different from static beam
TEM: a small spot size is formed on the sample surface with the condenser lenses. This probe is
scanned on the sample surface with scan coils, and in parallel, the signal is detected by an electron
detector, amplified and synchronously displayed on a cathode-ray tube (CRT) with the scan coils. The
detector can be a small disk on the column axis which detects only the transmitted beam (BF STEM
image) or a diffracted beam (DF STEM image), or can be an annular detector (a plate with a hole)
which detects all the diffracted beams except the transmitted one (ADF STEM). The resolution of the

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image is only limited by the spot size
(quality of the condenser lenses) since the objective lens is not used. In general, STEM images have
poorer resolution but better contrast than TEM images. ADF STEM with high angle (>50 mrad) can be
used to make chemical images1, but in general the contrast in ADF STEM images is determined by
the diffraction. This technique was used for the observation of the fine Si precipitates in the matrix of a
WFA/Al2O3 composite
3. Explain the working principle of Electron spin resonance spectroscopy with a block diagram. (16)
(May 2014)
ELECTRON SPIN RESONANCE ESR
SPECTROMETER Introduction
Electron spin resonance (ESR) spectroscopy has been used for over 50 years to study a variety of
paramagnetic species. Here, we will focus on the spectra of organic and organotransition metal
radicals and coordination complexes. Although ESR spectroscopy is supposed to be a mature field
with a fully developed theory there have been some surprises as organometallic problems have
explored new domains in ESR parameter space. We will start with a synopsis of the fundamentals of
ESR spectroscopy. For further details on the theory and practice of ESR spectroscopy, refer to one of
the excellent texts on ESR spectroscopy.
Operation of an ESR Spectrometer
Although many spectrometer designs have been produced over the years, the vast majority of
laboratory instruments are based on the simplified block diagram shown in Figure 3.1. Microwaves are
generated by the Klystron tube and the power level adjusted with the Attenuator. The Circulator
behaves like a traffic circle: microwaves entering from the Klystron are routed toward the Cavity
where the sample is mounted.

Microwaves reflected back from the cavity (less when power is being absorbed) are routed to the
diode detector, and any power reflected from the diode is absorbed completely by the Load. The diode
is mounted along the E-vector of the plane-polarized microwaves and thus produces a current
proportional to the microwave power reflected from the cavity. Thus, in principle, the absorption of
microwaves by the sample could be detected by noting a decrease in current in the
microammeter. In practice, of course, such a d.c. measurement would be far too noisy to be useful.
The solution to the signal-to-noise ratio problem is to introduce small amplitude field modulation. An
oscillating magnetic field is super-imposed on the d.c. field by means of small coils, usually built into
the cavity walls. When the field is in the vicinity of a resonance line, it is swept back and forth through
part of the line, leading to an a.c. component in the diode current. This a.c. component is amplified
using a frequency selective amplifier, thus eliminating a great deal of noise. The
modulation amplitude is normally less than the line width. Thus the detected a.c. signal is proportional
to the change in sample absorption. As shown in Figure 3.2, this amounts to detection of the first
derivative of the absorption curve

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The microwave-generating klystron tube requires a bit of explanation. A schematic


drawing of the klystron is shown in Figure 3.3. There are three electrodes: a heated cathode from
which electrons are emitted, an anode to collect the electrons, and a highly negative reflector electrode
which sends those electrons which pass through a hole in the anode back to the anode.

The motion of the charged electrons from the hole in the anode to the reflector and back to the anode
generates a oscillating electric field and thus electromagnetic radiation. The transit time from the hole
to the reflector and back again corresponds to the period of oscillation (1/n). Thus the microwave
frequency can be tuned (over a small range) by adjusting the physical distance between the anode and
the reflector or by adjusting the reflector voltage. In practice, both methods are used: the metal tube is
distorted mechanically to adjust the distance (a coarse
frequency adjustment) and the reflector voltage is adjusted as a fine control.

The sample is mounted in the microwave cavity, shown in Figure 3.4. The cavity is a rectangular
metal box, exactly one wavelength in length. An X-band cavity has
dimensions of about 1 2 3 cm. The electric and magnetic fields of the standing
wave are shown in the figure. Note that the sample is mounted in the electric field nodal plane, but at a
maximum in the magnetic field.

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4. What are the basic components of a Mass Spectrometer? Explain the functions of each component.
(16) (May 2014, May 2015, Nov/Dec 2016)

MASS SPECTROPHOTOMETRY
Over the past decade, mass spectrometry has undergone tremendous technological improvements
allowing for its application to proteins, peptides, carbohydrates, DNA, drugs, and many other
biologically relevant molecules. A mass spectrometer determines the mass of a molecule by measuring
the mass-to-charge ratio (m/z) of its ion. Ions are generated by inducing either the loss or gain of a
charge from a neutral species. Once formed, ions are electrostatically directed into a mass analyzer
where they are separated according to m/z and finally detected. The result of molecular ionization, ion
separation, and ion detection is a spectrum that can provide molecular mass and even structural
information.

According to the Lorentz force and Newton's second law in electromagnetic field, we have the
following equation.

So for different mass/charge ratio, the ions in the electromagnetic field can have different
trajectories. Solving the above two equations, we get the mass to charge ratio should satisfy:

Therefore, as different molecules have different mass to charge ratio, they follow different trajectories
in the electromagnetic field. Their trajectories should also depend on their initial speed and the
directions of the fields, but these can be calculated as well.
There are many types of mass analyzers, using either static or dynamic fields, and magnetic or electric
fields, but all operate according to this same law. Each analyzer type has its strengths and weaknesses.
Many mass spectrometers use two or more mass analyzers for tandem mass spectrometry (MS/MS). In
addition to the more common mass analyzers listed below, there are other less common ones designed
for special situations.
For the Time-of-flight (TOF) analyzer, it's just an electric field accelerating the ions through the same
potential.By measuring the time they take to reach the detector, when the charge is known, the mass
can be determined. If the particles all have the same charge, then their kinetic energies will be
identical, and their velocities will depend only on their masses. Lighter ions will reach the detector
first.

TIME OF FLIGHT MASS SPECTROMETERS


The time of flight mass spectrometer
In a time of flight mass spectrometer, ions of different mass/charge ratio are separated by the
difference in time they take to travel over an identical path from the ion source to the collector. This

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requires the starting time to be well defined .Therefore, ions are either formed by a pulsed ionisation
method or various kinds of rapid electric field switching are used as gate to release the ions from the
ion source in a very short time.in a pulsed mass spectrometer, ion packets of a few microseconds
duration are emitted at intervals of a few milliseconds from a voltage source. The ions traverse an
evacuated tube called the drift tube to reach the detector .The detector is sensitised for a brief, the
accurate measurement of the time between activating the source and sensitising the detector gives
information regarding the mass of the ions. The signal from the ions reaching the detector is
amplified and applied to the vertical deflection plates of the oscilloscope. The horizontal axis
deflection of the oscilloscope commences as the ion packets start out. This produces a mass spectrum
on the screen of the oscilloscope.
The device, thus gives a mass spectrum in a very short time. The essential parts of a time of flight

and suitable electronic circuitry for translating the time dependant arrival of ions of different velocities
into a time base that is related to mass number

If L is the length of the drift tube in centimetres and t is transit time in microseconds, for singly
charged ions of mass m and constant energy Ve, then
T=L[m/2Ve]1/2
=L[m/e1/2 V]1/2
m/e=2V/L2t2
if the detector is sensitized for a period t at a time t, the resolution m/m 2t/t
the above equation shows that the time resolution will increase with increased drift tube length and
will decrease with increasing accelerating voltage.
QUADRUPOLE MASS SPECTROMETERS
Quadrupole type mass spectrometer are simple in construction and light weight as compared to sector
type mass spetrometers,high speed electronic scanning and low cost .a quadrupole mass spectrometer
consists of an ion source, a quadrupole mass filter and a lens system to focus the ions into the
quadrupole filter. The arrangement consisting of four cylinder rod shaped electrodes which provide a
potential field distribution, periodic in time and symmetric with respect to the axis which will transmit
a select mass group and cause ions of improper mass to be deflected away from the axis. This mass
selection scheme uses a combination of dc potential plus a radio-frequency (rf) potential. By proper
selection of potentials and frequency, an ion of the desired mass can be made to pass through the
system, while unwanted masses will be collected on one of the electrodes.

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In practice, opposite electrodes of the filters are connected together, and to one pair a potential
(t)=u+vcos(2ft)is applied ,where u is a dc voltage and v is the peak amplitude of rf voltage at a
frequency f.to the other pair of electrodes ,the same potential but of the opposite sign is applied. Ions
emitted out of the ion source and focused into the quadrupole filter are made to undergo transverse
motion by the rf and dc field perpendicular to the Z-axis.

5. Draw the block diagram of an NMR spectrometer. Explain the function of each part and explain how it
is used to obtain NMR spectra. How are these spectra useful? (May 2013, May 2015, May/ June 2016)
Nuclear magnetic resonance (NMR) is a physical phenomenon in which magnetic nuclei
in a magnetic field absorb and re-emit electromagnetic radiation.
This energy is at a specific resonance frequency which depends on the strength of the magnetic
field and the magnetic properties of the isotope of the atoms
It is the study of absorption of radio frequency radiation by nuclei in a magnetic field. For a
particular nucleus a NMR absorption spectrum may consists of one to several group of
absorption lines in the radio frequency region of the electromagnetic spectrum.
They indicate the chemical nature of the nucleus and spatial positions of neighbouring nuclei.
Most powerful techniques for chemical analysis.
NMR spectrometer provides an accurate and non-destructive method of determining structure
in liquids and soluble chemical compounds.
In NMR spectroscopy the characteristic absorption of energy by certain spinning nuclei in a
strong magnetic field, when irradiated by second and weaker field perpendicular to it permits
the identification of atomic configurations in molecules.
Absorption of energy occurs when these nuclei undergo transition from one alignment in the
applied field to another one.
The energy absorption can be related to the magnetic dipolar nature of spinning nuclei.

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6. What are Mass spectrometers? Describe magnetic deflection mass spectrometers. (May 2013)
The heated tungsten filament produces an electron beam, which passes between plates A
and B.
A difference in electrical potential between A and b pulls ions out of the beam, so that they
pass through the slit in B into the region between B and c. the potential difference between b and c.

is adjustable from 0 up to several thousand volts. The ion beam than enters the space between two
trapezoid shaped magnet poles where it is deflected through an angle of 60, 90,120 or 180.
When ions of mass m and charge e pass through an accelerating electric field, they would attain a
velocity v which can be expressed in terms of the accelerating voltage v as follows 1/2mv2=eV
------- (1)
Where 1/2 mv2 is the kinetic energy of the ion as it leaves the electric field
From equation (1), v can be written as
V= [2ev/m]1/2 ------- (2)

If the ions next enter a magnetic field of constant intensity h, which is applied at right angles
to their direction of motion the ions would be diverted into circular orbits. From physics it is known
that when acceleration is applied perpendicular to the direction of ion motion, the objects velocity
remains constant, but the object travels in a circular path. Therefore, the magnetic sector follows an
arc. The radius and angle of the arc vary with different optical designs. Equating the centripetal and
the centrifugal forces:
mv2/r=hev ------- (3)

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The radius of curvature of the trajectory is given by:
r=mv/eh ------- (4)
Substituting for v from equation (2) r=
[2ev/m]1/2*m/eh
= [2ev/m*m2/e2h2 ]1/2
= [2v/h2*m/e]1/2
This equation shows that the radius of the orbit is a function of the mass/charge ratio of the particles
Applications of mass spectrometer:
In addition to general inorganic analysis and trace analysis in inorganic chemistry, rocket-borne mass
Spectrometers now analyze the upper atmosphere. Perhaps the most impressive experimental
achievement in mass spectroscopy is the successful mass spectrometric analysis carried out on the
surface of mars. Process monitor mass spectrometers, with the idea of direct control of industrial
process have been constructed.
An important requirement for studying respiratory physiology in routine clinical investigations is the
continuous analysis of the gas flow in a patients breathing cycle. A mass spectrometer would be
capable of analyzing gases in the mass range 15 to 50, whichcovers the full range of respiratory gases
from water to carbon dioxide.
In the early 1950, mass spectrometers were used for the quantitative analysis of mixtures of the
lighter hydrocarbons containing molecules with up to six or seven carbon atoms. The mass spectra of
the various components in a mixture were taken to be linearly additive, and the analysis was worked
out by setting.

7. Explain time of flight and quadrupole mass spectrometers.

The time of flight mass spectrometer


In a time of flight mass spectrometer, ions of different mass/charge ratio are separated by the
difference in time they take to travel over an identical path from the ion source to the collector. This
requires the starting time to be well defined .Therefore, ions are either formed by a pulsed ionisation
method or various kinds of rapid electric field switching are used as gate to release the ions from the
ion source in a very short time.in a pulsed mass spectrometer, ion packets of a few microseconds
duration are emitted at intervals of a few milliseconds from a voltage source. The ions traverse an
evacuated tube called the drift tube to reach the detector .The detector is sensitised for a brief, the
accurate measurement of the time between activating the source and sensitising the detector gives
information regarding the mass of the ions. The signal from the ions reaching the detector is
plates of the oscilloscope. The horizontal axis deflection of the oscilloscope commences as the ion
packets start out. This produces a mass spectrum on the screen of the oscilloscope. The device, thus
gives a mass spectrum in a very short time. The essential parts of a time of flight instrument are

An electron gun for the production of ions


A grid system for accelerating ions to uniform velocities in a pulsed mode
An evacuated tube called the drift tube and
An ion detector and suitable electronic circuitry for translating the time dependant arrival of
ions of different velocities into a time base that is related to mass number
If L is the length of the drift tube in centimetres and t is transit time in microseconds, for singly
charged ions of mass m and constant energy Ve, then T=L[m/2Ve]1/2
=L[m/e1/]1/2
m/e=2V/L2t2
if the detector is sensitized for a period t at a time t, the resolution m/m 2t/t
the above equation shows that the time resolution will increase with increased drift tube length and
will decrease with increasing accelerating voltage.
The current produced by the ions arriving at the collector may have a very short duration,
which necessitates the use of a wide band amplifier. A specially designed magnetic electron multiplier
is used for this purpose. this multiplier uses a strip of semiconducting material for the multiplying
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surface instead of dynodes. A gain of 10 is attained with a dark current of only 310 -21 A .the ions
7
leaving the ion source of a time of flight mass spectrometer have neither exactly the same starting
times nor exactly the same kinetic energies. Various time of flight mass spectrometer designs have
been developed to compensate for these differences .one such design makes use of a device known as
reflectron. The reflectron is an ion optic device in which ions in a time of flight mass spectrometer
pass through a mirror and their flight is reversed.
The linear reflectron allows ions with greater kinetic energies to penetrate deeper into the reflectron
than ions with smaller kinetic energies. The ions that penetrate deeper will take longer to return to the
detector .if a packet of ions of a given mass to charge ratio contains ions with varying kinetic energies,
then the reflection will decrease the spread in the ion flight times, and therefore improve the resolution
of the time of flight mass spectrometer. The arrangement is shown below.

8. Describe the working of double beam mass spectrometer and give its
applications. Refer Q.No : 4

9. Explain the instrumentation and applications of Scanning Electron Microscope (SEM) and
Transmission Electron Microscope (TEM). (Nov/Dec 2016, May/ June 2016)
Refer Q.No : 1 and 2
10. Describe how various samples are analyzed using NMR spectrometer with neat
diagram. Refer Q.No : 5
11. Explain the construction and working principle of Electron Spin Resonance (ESR) spectrometer with
neat diagram. ( Dec 2015, Nov/Dec 2016)
Refer Q.No : 3
12. Explain the construction and working of Radio frequency spectrometer with neat diagram.
Refer Q.No : 3
13. Explain the various components of a mass
spectrometer. Refer Q.No : 4
14. Explain in detail the construction and working principle of single focusing mass spectrophotometers.
(Nov 2013)
Refer Q.No : 4
15. In NMR spectroscopy mention the advantages of using a magnet with as great a field strength as
possible. Also explain the difference between a continuous wave and fourier transform NMR.
Refer Q.No : 5
16. With neat sketch explain the construction and working principle of mass spectrometer.
Refer Q.No : 4
17. What are the basic components of electron Spectroscopy? Also explain the working principle of
Electron spectroscopy with a neat block diagram. (Nov 2013)
Refer Q.No : 3

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18. Explain the construction and working principle of Electron Spin Resonance (ESR) spectrometer with
neat diagram.( Dec 2015)
Electron spin resonance is the study of magnetic dipoles of electronic origin by applying fixed
microwave frequencies to a sample residing in a varying magnetic field. Its also known as Electron
Paramagnetic Resonance (EPR) spectroscopy.
Below shown is the block diagram of ESR spectrometer.

The sample is irradiated by microwave energy from the klystron in the microwave bridge. A klystron
normally operated at 9.5GHz, generates a microwave field. The magnetic field at the sample is
modulated at 100 KHz. The klystron output passes through an isolator, a power leveler and a
directional coupler. The filed is applied to the resonant cavity, which is connected to one arm of the
microwave bridge. During the magnetic field scan, when field intensity reaches the value required to
induce electron spin resonance in sample, a change occurs in the amount of microwave energy
absorbed by the sample. This causes a change in the microwave energy reflected from the cavity. The
reflected microwave energy, which is modulated at the field modulation frequency, is directed to the
detector crystal. The detector is usually a silicon tungsten crystal rectifier. After detection at the
crystal, the resulting 100 kHz, which contains the ESR information, is amplified in the pre-amplifier
circuit and applied to the receiver section of the 100 kHz modulation unit. The amplified signal is
phase-detected to obtain the spectrum, which appears as a deflection on the Y-axis of the recorder.
The field scan potentiometer is linked mechanically to the recorder X-axis. The X-axis is calibrated in
gauss. The ESR signal may also be applied to the oscilloscope for visual display. The field controller
accurately controls the magnetic field to the desired set value. The important operating parameters of a
typical ESR spectrometer (Figure 11.6) are: Operating frequency 8.8 to 9.6 GHz Sensitivity 5 x 10"
H Spins Modulation frequencies 1000 kHz, 270 Hz, 35 Hz Modulation amplitude range 5 mG to
40 G in steps rf power to cavity 200 mW calibrated over full frequency range 15 mW
a)The magnet and Magnetic field controller.

The magnet used in the ESR spectrometers is usually of the electromagnet type. It provides a
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homogeneous magnetic field, which can be varied from 200 mgauss to 20 kgauss, calibrated in steps.
Stability of 1 part in 106 is satisfactory for adequate resolution of ESR spectra. The magnetic field
controller provides direct control and regulation of the magnetic field in the air gap. It is an ac carrier type
servo system that accurately controls the magnetic field. Figure 11.7 shows the schematic diagram of the
magnetic field controller in a spectrometer. The driver amplifier supplies a 30 mA rms, 1230 Hz exciting
current through the field set reference resistor (R), the Hall effect magnetic field sensor element and the
primary of the scan voltage transformer (T4). A field reference voltage is developed across R. Similarly, a
field scan reference voltage is developed across the constant input impedance of scan range attenuator by
the transformed reference current in the secondary winding of T4. The output of the summation circuit (i.e.
input to the error amplifier) is the algebraic sum of the output of the Hall-effect field sensor (transformed
by T5) added to the field set and field scan voltage. The sum of these three voltages is amplified by the
error amplifier and applied to the input of the phase-sensitive detector. When the output of the Hall-effect
field sensor is less than the sum of the field set and field scan voltages, the output of the phase-sensitive
detector is of a polarity which turns on the magnet power supply, to increase the magnet current and thus
the magnetic field. In a similar manner, when the output of the Hall-effect field sensor exceeds the sum of
the field set and field scan voltages, the current through the magnet winding is decreased and the magnetic
field is decreased. The field scan potentiometer is centre-tapped and the wiper is driven by the recorder's
hori-zontal axis-drive mechanism. The voltage output of the scan potentiometer acts to oppose the field set
voltage, when the recorder pen is in the left-half of its operating range, but adds to the field set voltage,
when the recorder pen is moved into the right half of the recorder chart. The 1230 Hz oscillator is a stable

Wein bridge oscillator, whose output level is internally regulated. The error amplifier is a high gain
stable amplifier with a pass-band centred near 1230 Hz. It has a phase shifting potentiometer, which
permits the phase to be adjusted, so that the signal component of the amplified error signal is exactly
in-phase and the spurious quadra tore component of the amplified error signal is exactly out-of-phase,
so that the latter is com-pletely rejected by the phase sensitive detector. The phase-sensitive detector
converts the amplified 1230 Hz output signal of the summation circuit into dc control voltages to
drive the magnet power supply. The Hall probe is maintained at a constant temperature. The
temperature control is main-tained by controlling current through a hater, sensed by a thermistor.
The magnet power supply provides controlled dc current to the low impedance electromag-net. The
magnet current is supplied by the main transformer and rectifiers. The magnets used could be of 9, 12
or 15 inches. They are generally floor-mounted and require 2.5 to 22.5 kW power supplies to drive
them. The magnets are equipped with ring-shim pole caps designed for maximum field homogeneity.
A 3-inch clearance (air gap) provides ease of access. The pole caps are made compatible with tapered
ring-shim tips, which reduce the air gap to 1.75 inch to achieve optimum field performance when
operating at 35 GHz. In order to improve magnet performance, the heat generated must be dissipated.
To do so, coolant lines are run around the magnet, magnet power supply and microwave bridge. If the
temperature of the untreated water is below the dew point, condensation may occur in the magnet
power supply. In this event, the untreated water can be run through the magnet first and then through
the power supply. Also, the coolant water flow should be correct for magnet system in use. This is
ensured by checking the difference between inlet and outlet pressure if the difference is more than that
specified for the magnet system, the cooling system may be clogged. If the ambient room-air
temperature or magnet cooling water temperature changes, the setting of the Hall probe temperature
should be examined to ensure that the temperature control system is regulating, i.e. it is not saturated
or cut off. This is particularly important in those installations wherein large annual variations in room
or water temperatures take place. Noise and jitter can be induced into the field control system by
transients on the power lines, that are coupled into the Hall probe circuit. As far as possible, the Hall
probe cable should be isolated from power lines connecting the console and the magnet power supply.
It is often difficult to detect or troubleshoot instabilities of the magnetic field, as normally no
convenient external field measuring device is available to monitor the magnetic field with sufficient
stability or accuracy. However, the manufacturers of instruments suggest several test techniques that
may be employed with easily obtainable auxiliary test equipment. If the magnet current rises beyond
normal, the Hall sensor probe may be improperly phased. This is corrected by removing power from

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the system and reversing power leads to magnet coils.
b)The Micro Wave Bridge
The microwave bridge detects the ESR signal reflected from the sample cavity. The bridge contains
microwave circuitry, a klystron power supply, a pre amplifier and an automatic frequency control circuit
(Figure 11.8). The klystron generates microwave energy at 93 GHz, which is used to irradiate the
sample. The klystron output is applied to an isolator which allows signal flow only in one direction. The
klystron is water-cooled through its mounting flange. 35 GHz systems are also avail-able. The output of
the isolator is given to a four port circulator, wherein the microwave power is directed to the sample
cavity. The circulator then directs the reflected power from the sample cavity back to the microwave
detector. The crystal do bias current is indicated by the detector level meter.
When all the power is absorbed and none is reflected, the detector receives no signal. At resonance,
the sample absorbs microwave energy and the unbalances the impedance of the cavity. The absorption
in the sample is detected as a dip in the output of the crystal detector. The crystal output containing
the ESR signal and the 70 kHz AFC (automatic frequency control) signal is amplified by the pre-
amplifier. It is then coupled to the receiver and to the AFC amplifier. The automatic frequency
control circuit contains a 70 kHz crystal-controlled oscillator, which generates an AFC carrier and the
AFC phase detector reference voltage. The AFC carrier is
superimposed on the klystron reflector voltage, which results in a 70 kHz frequency modulation of the
klystron output. The frequency modulated rf (radio-frequency) is applied to the sample cavity,
resulting in an fm (frequency modulation) to am (amplitude modula-tion) conversion. The resultant
am modulated microwave power is detected and amplified in a preamplifier, followed by a 70 kHz
tuned amplifier. The output of this amplifier is phase-detected, amplified, filtered and applied to the
klystron reflector tracking network. Another parallel output is given to an integrator, which quickly
returns the phase detector dc output to zero. This arrangement permits a very accurate lock, over a
range of at least 15V reflector voltage correction. Noise elimination is an important factor in
enhancing the sensitivity of plotting the ESR spec-trum. Most of the noise is eliminated in the phase-
sensitive detector, because only that pad of the noise which is at the same frequency and in-phase with
the reference signal, is allowed to pass through it. The noise is further eliminated by using a long-time
constant filter, which averages out noise at frequencies greater than the reciprocal of its time constant.
A better method of noise elimination is through continuous averaging by a computer, which employs a
multichannel pulse-height analyser. This technique of using a computer for average transients
efficiently removes both low and high frequency noise. In order to achieve advantages of higher
frequency operation such as increased sensitivity for small samples, minimization of second order
shifts, increased resolution for powder samples with different g-values, the klystron frequency is set at
35 GHz instead of 9.5 GHz. The higher fre-quency also permits the observation of transitions that
require higher energy. The klystron requires +650V for the klystron beam and up to 400 V for the
klystron reflector. The supply voltages are obtained from the 20 V dc inputs in inverter circuits,
working at a frequency of 35 kHz. This frequency is obtained by dividing the 70 kHz AFC modulation
frequency by 2. The klystron body or tuning shaft should not be touched when the bridge is in tune or
in the operate position because the klystron body is at +650 V when operating. It is insulated from
ground by an insulating gasket placed between the klystron flange and the water cooling flange.

c)Modulation Unit
The 100 kHz modulation unit acts as a transmitter and receiver. The transmitter provides the
power to drive the cavity modulation coils. The receiver processes the ESR signal from the bridge pre-
amplifier and converts it into a do voltage for application to the recorder Y-axis. Figure 11.9 shows
the block diagram of the 100 kHz modulation unit. The 100 kHz oscillator is crystal-controlled. Its
output is attenuated by the modulation amplitude control and is applied to the output modulation
amplifier. The gain of the ampli-fier is controllable. The 100 kHz modulation signal is transmitted to
the cavity modulation coils.
The 100 kHz ESR signal from the detector crystal in the bridge is filtered and amplified in the 100
kHz receiver. The receiver gain control can be set in 1 dB steps. The amplified 100 kHz ESR
signal is applied to the input of the phase detector. The amplitude of the dc phase detector output is

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proportional to the amplitude of the 100 kHz signal input. A low-pass filter removes any residual 100
kHz or harmonics from the phase-detected signal. Additional noise filtering is provided by an RC
network, which provides a variable time constant. This is followed by a buffer amplifier, which
provides low output impedance stage connection to the recorder. Modulation may also be carried out
at low frequencies like 10 kHz, 1 kHz, 270 Hz or 35 Hz. The low frequency module shown on the
block diagram serves to accomplish this function.
d) System Function Selector
The system function selector provides the selection of signal inputs to the recorder and oscilloscope
and cavity modulation to the cavity. When the oscilloscope monitor and the recorder input switches
are in the 100 kHz or low frequency position, the phase-detected ESR signal passes from the receiver
to the oscilloscope and recorder respectively. In the external position, the recorder input may be
supplied from an external source and the oscilloscope may be fed an external signal. The cavity
modulation switch allows the operator to apply either 100 kHz or a low frequency modulation to the
cavity coils. If two cavities are used, a low frequency can be applied to one superimposed on the
klystron reflector voltage, which results in a 70 kHz frequency modulation of the klystron output. The
frequency modulated rf (radio-frequency) is applied to the sample cavity, resulting in an fm (frequency
modulation) to am (amplitude modula-tion) conversion. The resultant am modulated microwave power
is detected and amplified in a preamplifier, followed by a 70 kHz tuned amplifier. The output of this
amplifier is phase-detected, amplified, filtered and applied to the klystron reflector tracking network.
Another parallel output is given to an integrator, which quickly returns the phase detector dc output to
zero. This arrangement permits a very accurate lock, over a range of at least 15V reflector voltage
correction. Noise elimination is an important factor in enhancing the sensitivity of plotting the ESR
spec-trum Most of the noise is eliminated in the phase-sensitive detector, because only that pad of the
cavity and 100 kHz to the other cavity, or the two modulation signals may be added and applied
simultaneously to either cavity. The 100 kHz modulation and the 34 kHz sawtooth sweep are
combined in the modulation summation circuit. Two decade capacitors series resonate the cavity
modulation coils at 100 kHz for the most efficient energy transfer to the cavity. The ESR signal can be
observed on the oscilloscope with 100 kHz or 10 kHz modulation, but not with any of the other lower
frequencies.
11.3.5 Recorder
The recorder is used to produce a hard copy of the ESR spectrum, as a function of time on a 30 x 40
cm flat chart. By selecting the synchronized frequency to the stepper motor, the recording time can be
selected from a few seconds to several hours. Horizontal travel (X-axis) of the pen is achieved by a
pulsed stepper motor and belt-driven capstan, which moves the pen carriage horizontally. The vertical
(Y-axis) travel of the pen is controlled through a second system of pulleys and wire cable from a servo
motor. Y-axis movement results when the servo loop, of which motor is a part, is unbalanced by a
signal applied to the unbalanced by a signal applied to the recorder amplifier. The recorder amplifier
is turned on by the servo motor to balance the servo loop.
11.3.6 Oscilloscope
The oscilloscope permits direct visual observation of rapidly changing or decaying signals as well as
the optimization of instrument parameters. The oscilloscope supplies 34 Hz sawtooth modulation to
the sample cavity modulation coils and displays the resulting ESR spectrum. The amplitude of the
field modulation is kept adjustable and may be selected by sweep width control. Noise on the
displayed spectrum may be reduced by the addition of an input filter. The oscilloscope is also used to
display the klystron mode. It is accomplished by applying the same 34 Hz sawtooth modulation to the
klystron reflector and displaying the output of the microwave detector on the oscilloscope.
113.7 Sniffle Cavities
Signal intensity from a particular sample can vary greatly, depending upon the microwave cavity
configuration. This is important when signal-to-noise poses a serious problem. The two main
effects arise from: (i) the difference between cylindrical (TE017) and rectangular cavities (TE012),
(ii) the effect of quartz such as Dewar inserts, on the microwave field distribution. The use of a
cylindrical cavity results in a net improvement over the rectangular cavity. Both types of cavities
are, however, transverse electric that the electric field

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Transverse electric modeslines are confined to the plane that is perpendicular to the longitudinal axis.
No such restriction exists for the magnetic field lines. The sample cavity should be so designed that it
can be held in the volume of maximum homogeneity of the magnetic field. The cavities are available
for a wide variety of ESR signals, ranging from solids to liquids. They are designed to be compatible
with other accessories like aqueous solution sample cell, flow mixing chamber, electrolytic cell, liquid
nitrogen Dewar and the variable temperature accessory.
ESR studies involving comparative measurements are made by using a dual cavity. This cavity does
this by allowing a reference sample of known magnetic resonance characteristics to be exposed to
microwave energy simultaneously with and in the same cavity as the sample being studied.
A rotating cavity is designed to facilitate ESR studies of crystal anisotropy. The cylindrical rotating
cavity has exterior modulation coils, which are free to turn with the rotating electromagnet and thus
provide a modulation field, which is always parallel to the dc magnetic field. The sample tube remains
fixed in the cavity, which keeps the Q constant and eliminates the need for re-tuning after rotation.
11.3.8 Sample Cells
Aqueous solution sample cells are specially designed to carry out FSR studies in lossy or aqueous
solutions. Further, as optical irradiation experiments are often performed with these cells, the quartz
is selected to give a maximum optical transmission in the UV-visible region of the spec-trum. Special
types of flow cells, tissue cells and electrolytic sample cells are also used when
required. Generally, the standard sample tube is made of high purity quartz and measures 3 mm ID and
4 mm OD. Glass is not used because it contains traces of FE3'. For maximum sensitivity, the tube may
be filled to a height of 2.5 cm for a rectangular cavity and 5 cm for a cylindrical cavity. The single
crystal method of ESR analysis requires a large number of measurements with precise orientations of
the sample. Such measurements are often difficult, because of the small size of microwave cavities.
Some goniometers for ESR studies have been described in literature, but either they only permit 360*
rotation about a single axis, or the angular variation of the signal/noise ratio (modulation coils not
aligned with H.), prevents the spectrum from being recorded for some orientations. All sample tubes
should be cleaned before being inserted into the cavity. Only standard ESR sample tubes should be
used to prevent breakage and to minimize contamination signals. Sources of contamination such as
cigarette ashes or smoke should be kept away from the immediate vicinity of the cavity, whether the
cavity is installed in the air gap or not. Loose magnetic materials must be kept away from the gap of
the magnet to prevent damage to the cavity, when it is in the air gap. Watches should be kept away
from the magnet gap. Sample materials or solvents (water-acetone) with high dielectric loss require
flat cells. For materials with low dielectric, a regular 3 mm sample tube is used.

19. With neat sketch explain the construction and working principle of mass spectrometer. (Dec 2015)
Components of a Mass spectrometer
The following five units are common to most mass spectrometer instruments: the inlet
sample system, the ion source, the electrostatic accelerating system, the detector, amplifier
and display system, and auxiliary equipment (pumping system).
9.4.1 The Inlet Sammie System
Gaseous Samples: The introduction of gases merely involves the transfer of the sample from a gas
bulb into the metering volume. The arrangement is a small glass manifold of known volume
attached to a mercury manometer. The pressure range is generally from 30 to 50 mm of Hg. The
gas sample is introduced into the mass spectrometer ion source through a leak of some kind.
Generally, the leak is a pin-hole in metal foil. Hogg (1969) explains the construction of a variable
leak inlet system used in high resolution mass measurement Liquid Samples: Liquid samples may
be introduced by a hypodermic needle and injected through a silicon rubber dam, or by a break-
off device which consists in touching a micropipette to a sintered glass disc under mercury. The
low pressure in the reservoir draws in the liquid and vaporizes it instantly. Solid Samples: Solid
samples can be vaporized to gaseous ions by instantaneous discharges with a power up to 100 kW
by using a radiofrequency (1 MHz) spark. Under these conditions, all constituents of the sample
are converted to gaseous form at an equal rate without regard to their vapour pressure, thereby

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EI6501 Analytical Instruments Dept. of EIE and ICE 2016-2017
eliminating the possibility of preferential vaporization
9.4.2 Ion Sources
Following the leak is the ionization chamber, which is maintained at a low pressure (10 to 10-7
mm Hg) and at a temperature of 200C. The electron gun is located perpendicular to the incoming
gas stream. Electrons are emitted from filament (Figure 9.11) normally of carbonized tungsten, but
for special purposes, tantalum or oxide coated filaments may be used. They are drawn off by a pair
of positively charged slits, through which they pass into the body of the chamber. The potential
present in the slits controls the electron emission and the energy of the electrons. An electric field
applied between these slits accelerates the electrons, which on subsequent collisions with
molecules of the passing gas stream, produce ionization and fragmentation. In order to obtain a
mass spectrum, the electric field is kept between 50 and 70 V. The electron beam is usually
collimated by a magnetic field, which is confined to the ionization region.
9.4.2.1 ICP as Ion Source
ICP (inductively coupled plasma) has been shown to be a potentially effective ion source (Houk,
1986). ICP hardware is designed to generate plasma, which is a gas in which atoms are present in
an ionized state. The basic set-up of an ICP consists of three concentric tubes, most often made of
silica. These tubes, termed outer loop, intermediate loop, and inner loop, collectively make up the
TCP of the Torch. The torch is situated within a water-cooled coil of a ICP Torch radiofrequency
(rf) generator. As flowing gases are introduced into the torch, the rf field is activated and the gas
in the coil region is made electrically conductive. This sequence of events forms the plasma. The
formation of the plasma is dependent upon an adequate magnetic field strength and the pattern of
the gas streams follows a particular rotationally symmetrically pattern. The plasma is maintained
by inductive heating of the flowing gases. The induction of a magnetic field generates high
frequency annular electric current within the conductor. The conductor, in turn, is heated as the
result of its ohmic resistance. In order to prevent possible short-circuiting as well as meltdown, the
plasma must be insulated from the rest of the instrument. Insulation is achieved by the concurrent
flow of gasses through the system. Three gases flow through the systemthe outer gas,
intermediate gas, and inner or carrier gas. The outer gas is typically argon or nitrogen. The outer
gas has been demonstrated to serve several purposes including maintaining the plasma, stabilizing
the position of the plasma, and thermally isolating the plasma from the outer tube. Argon is
commonly used for both the intermediate gas and inner or carrier gas. The purpose of the carrier
gas is to convey the sample to the plasma. However, the ICP operates at atmospheric pressure,
whereas mass spectrometer (MS) requires an operating pressure 10 torr for resolution and
detection of ions. The solution acts as an interface between the two. The conventional method of
sample introduction for inductively coupled plasma mass spec-troscopy is by aspiration, via a
nebulizer, into a spray chamber. A small fraction of the resulting aerosol is swept by argon into the
torch. Approximately 1 ml of sample is required per analytical run, about 99 per cent of which is
wasted. Recently, low cost, low uptake rate, high efficiency nebulizers have been employed to
combat this problem. The high efficiency nebulizer operates more efficiently at 10-2001/min. The
detec-tion limits and precision obtained with the high efficiency nebulizer are superior to
conventional nebulizers. Figure 9.12 shows the arrangement presently used for extracting ions
from the ICP. The plasma flows around the tip of a water-cooled metal cone called the sampler.
This cone has a circular orifice of 0.5-1.00 mm diameter drilled into its tip. Gas from the ICP
enters this orifice from a cross-sectional area of diameter about three times the orifice diameter, so
that a 1.5-3 mm wide section of the axial channel is sampled. Most of the gas flow is evacuated by
a mechanical pump that maintains a pressure of the order of 1 torr. The central orifice of a conical
skimmer is located behind the sampler in such a way that the sampled beam is transmitted into a
second vacuum chamber. The pressure here is low enough, and the mean free path long enough
for ion lenses to coiled, focus and transmit the ions to the mass analyser. The formation of singly
charged ions is very efficient in the ICP. Some 54 elements are ex-pected to be ionized with an
efficiency of 90 per cent or more. Even-metalloid or non-metallic elements such as P and As, for
which ionic emission lines are either absent or not prominent, are ionized with reasonable
efficiency. Therefore, a combination of the ICP with MS shows promise of extending the

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sensitivity and selectivity of MS to elemental analysis of solutions.
9.4.2.2 Glow Discharge Ion Source
In recent years, the glow discharge has been developed as a low-energy alternative ion source,
which, coupled with a quadrupole mass filter, serves as a simple and inexpensive mass spectrom-
eter. Basically, the glow discharge is a simple two electrode device, filled with a rare gas to about
0.1-10.0 torr. A few hundred volts applied across the electrodes cause breakdown of the gas and
formation of the ions. Figure 9.13 shows the basic components and discharge regions. The sample
to be analyzed serves as the cathode, whereas the anode material is not particularly critical The
sample is atomized into the discharge by a process known as sputtering. The glow dis-charge not
only atomizes the solid sample, but also provides the means by which these atoms are ionized.
Sputtered atoms diffuse into the negative glow, which contains energetic electrons, ions and
metastable atoms. We are basically concerned with the ionization of the sputtered atoms, rather
than the bulk ionization process, by which the discharge sustains itself. The glow discharge is
adaptable to many discharge types and ion source configurations. Some-times, the sample type
itself dictates a certain cathode geometry, whereas other source models arise from more
fundamental considerations. Glow discharge ion sources have been operated in the dc, pulsed-dc
and rf modes. The glow discharge source presents a flowing gas load to the mass spectrometer,
requiring differential pumping to interface the 1-torr source with the 10 6 to 10 7 torr analyzer
chamber. Most of the reported work in glow discharge mass spectrometers has been carried out
using quadrupole analyzer. This analyzer makes it possible to design instruments with low
sampling voltages and high ion transmission, particularly at low mass ranges. The glow discharge
possibly ionizes only a small percentage of the sputtered sample atoms. Coupling the output of a
tunable laser to the glow discharge plasma permits very selective ionization of a given element.
The focused laser beam is capable of close to 100 per cent ionization efficiency. In this
arrangement, the glow discharge functions primarily as an atomization source and the laser as the
means of ionization. Harrison et al. (1986) have reviewed the developments in glow discharge
mass spectrometry.
9.4.2.3 Other Ionization Methods
The various other ionization methods in use are described as follows. Electron Impact: An electron
impact source uses an electron beam, usually generated from a tungsten filament, to ionize gas-phase
atoms or molecules. An electron from the beam knocks an electron off of analyze atoms or molecules
to create ions. Electro-spray Ionization: This source consists of a very fine needle and a series of
skimmers. A sample solution is sprayed into the source chamber to form droplets. The droplets carry
charge when they exit the capillary. As the solvent evaporates, the droplets disappear leaving highly
charged analyze molecules. The method is particularly useful for large biological molecules that are
difficult to vaporize or ionize. Fast-atom Bombardment: In this method, a high energy beam of natural
atoms, typically Xe or Ar, strikes a solid or low vapour-pressure liquid sample causing desorption and
ionization. It is used for large biological molecules that are difficult to get into the gas phase. The
sample is usually dispersed in a matrix such as glycerol. The method causes little fragmentation and
usually gives a large molecular ion peak, making it useful for molecular weight determination. The
atomic beam is produced by accelerating ions from an ion source through a charge-exchange cell. The
ions pick up an electron in collision with neutral atoms to form a beam of high-energy atoms. Field
Ionization: Molecules can lose an electron when placed in a very high electric field. High fields can be
created in an ion source by applying a high voltage between a cathode and an anode called 'a field
emitter'. A field emitter consists of a wire covered with microscopic carbon den-drites, which greatly
amplify the effective field at the carbon points. Laser Ionization: A laser pulse ablates material from
the surface of a sample, and creates a micro-plasma that ionizes some of the sample constituents. The
laser pulse accomplishes both vaporiza-tion and ionization of the sample. Matrix-assisted Laser
Desorption Ionization: This is a laser-based method of vaporizing and ionizing large biological
molecules such as proteins or DNA fragments (Cotter, 1999). The biological molecules are dispersed
in a solid matrix such as nicotinic acid or dihydroxybenzoic acid. A UV laser pulse ablates the matrix
which carries some of the large molecules into the gas phase in an ionized form so they can be
extracted

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into a mass spectrometer. Thermal Ionization: Thermal ionization is used for elemental or
refractory materials. The sample is deposited on a metal ribbon, such as Pt or Re, and an electric
current heats the metal to a high temperature. The ribbon is often coated with graphite to provide
a reducing effect.
9.4.3 Electrostatic Accelerating System
Positive ions, which are separated from electrons by a weak electric field, are accelerated in a
strong electrostatic field between the first and second accelerating slits. Voltages of the order of
400-4000 volts accelerate the ions to their final velocities of up to 150,000 miles/s and they acquire
a kinetic energy of a few thousand electron volts. Such a relatively high kinetic energy is imparted
to the ions to produce an almost mono-energetic beam, when it finally emerges out of the final
accelerating slit, which is approximately 0.076 mm in width. The electrostatic voltages are highly
stabilized to an accuracy of better than 0.01 per cent.
9.4.4 Ion Detectors and Recording of Mass Soectrograun
Faraday cup: The ion beam passing through the exit slit of the analyzer tube is normally collected
in a cylinder (Faraday cage), which is connected to the grid of an electrometer tube, whose output
is in turn amplified. The use of an electrometer tube is necessitated because of an extremely low
magnitude of the ion current (104 to 104 A). Vibrating electrometers have also been used in order
to convert the dc current output into an ac signal. The amplified ion current is recorded as a
function of the ratio m/e on an oscillograph or a pen-and-ink strip chart recorder. Since a Faraday
cup can only be used in an analog mode, it is less sensitive than other detectors that are capable of
operating in pulse-counting mode. Channeltron: A channeltron is a horn-shaped continuous
dynode structure that is coated on the inside with an electron-emissive material. An ion striking the
channeltron creates secondary electrons that have an avalanche effect to create more secondary
electrons and finally a current pulse. Electron multiplier tube: Electron multiplier tubes are similar
in design to photomultiplier tubes. They consist of a series of biased dynodes that eject secondary
electrons when they are struck by an ion. They, therefore, multiply the ion current and can be used
in analog or digi-tal mode. Micro-channel plate: A micro-channel plate Inddent Ion consists of an
array of glass capillaries (10-25 micro meter inner diameter) that are coated on the inside with an
electron-emissive material. The capillaries are biased at a high voltage and like the channeltron, an
ion that strikes the inside wall of one of the plate capillaries creates an avalanche of second-ary
electrons. This cascading effect creates a gain of 103 to 10a and produces a current pulse at the
output. The schematic of a micro-channel plate is shown in Figure 9.14. The recorder must have a
provision for automatically recording peaks of widely varying am-plitudes. This is achieved by
using five separate recording channels with relative sensitivities of t 3, 10, 30 and 100,
respectively. This arrangement should enable to record the height of any peak within better than 1
per cent accuracy, over a range of magnitude of 1 to nearly 1000.
9.4.5 Vacuum System
In order to prevent undue scattering by collision of ions with residual gas molecules, the mass
spectrometer requires a good vacuum system. Generally, separate mercury or oil diffusion pumps
are employed in the source and analyzing regions of the spectrometers. Vacuum pump The two
important parameters of a vacuum pump are its lowest attainable pressure, and its pumping speed,
typically listed as litres per minute (1pm) or cubic feet per minute (cfm). The lowest attainable
pressure depends upon the design of the pump. The pumping speed of the different types of
pumps depends on the physical size of the pump. Pump Lowest Attainable Pressure Typical Use
Mechanical pump le - 10-1 torr Rouging or backing pump Diffusion pump 104 to Vacuum lines
Turbo molecular pump 104 torr High-vacuum systems
Mechanical pumps consist of an inlet, and exhaust with a one-easy valve, and art off-center
rotating piston in a cylindrical cavity. As the piston rotates, gas is pulled into the cavity, and forced
out through the exhaust port. The rotating piston has spring-loaded vanes to create a seal with the
cavity walls. This seal, and the exhaust port valve, are lubricated with a low-vapour-pressure oil. A
two-stage mechanical pump consists of two pumping cavities in series to achieve a lower vacuum
pressure. The accessories needed when mechanical pumps being, used are a mist titter or vent to
trap oil mist in the pump exhaust, and a trap to prevent oil vapour from backstreaming into the

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volume being evacuated. A diffusion pump consists of a bath of boiling oil that streams through a
jet-shaped volume. The oil entrains gas molecules and transports them in the direction of the oil
flow. A mechanical pump can then pump away the exhaust from the diffusion pump. A
turbomolecular pump or just turbo pump contains a turbine that is spinning at a very high rate of
revolution, typically tens of thousands revolutions per second. The turbine blades are spinning
faster than the average speed of gas atoms or molecules, so that any gas-phase species that enter
the turbo pump are physically forced out of the pump by the turbine blades. A mechanical pump
is required to maintain a low pressure and pump away the exhaust from a turbo pump. Many
analytical instruments including mass spectrometry, X-ray photoelectron and Auger electron
spectroscopies, and electron microscopy require a low pressure to operate. Special gad-gets or
instruments are required to measure such low level pressures.

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