Unit I Colorimeter and Spectrophotometers: 2. What Is Meant by Flame Emission Spectrometry?

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UNIT I
COLORIMETER AND SPECTROPHOTOMETERS
PART A
1.Radiant source
2.wavelength selector
3.photodetector
4.sample
2. What is meant by flame emission spectrometry?

When an analytical sample is introduced into the flame, the flame atomizes the
Sample and subsequently they are de-excited with the emission of atomic lines
corresponding to different elements present in the sample
3. State Beers law.

The intensity of a beam of monochromatic light decreases exponentially
with increase in concentration of the absorbing substance arithmetically.
4. What is monochromators?
Monochromators are the optical systems which provide better isolation of spectral energy
than the optical filters and are therefore preferred where it is required to isolate narrow bands of
radiant energy.
5. What is analytical instrumentation?

It is the field of science which deals with the technique by which the sample
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1. List any four elements used in spectrophotometers.
(chemical species) is analyzed quantitatively. Instruments used for analysis of
chemical sample are analytical instrument
6. What is electromagnetic radiation?

It is a type of energy that is transmitted through space at a speed of approximately 3x108
vacuum. It may be considered as discrete packets of energy called photons.

7. Define wave number.
It is defined as the number of waves per centimeter.
= 1/
8.
Draw the spectrum of EM regions.

Given in the notes.
9. State Lamberts law.

When a beam of light is allowed to pass through a transparent medium the rate
of decrease of intensity with the thickness of medium is directly proportional to
the intensity of light.
10. What is Absorption Spectroscopy?
The measurement method based on absorption of radiation of a substance is known as
absorption spectroscopy.
m/s. Such radiation doesnt require a medium of propagation and can readily travel through
PART B
1.
Explain with necessary diagrams the instrumentation involved in uv visible

spectrophotometer?
schematic diagram of a double-beam UV-Vis. spectrophotometer
Instruments for measuring the absorption of U.V. or visible radiation are made up of the
following components;
1. Sources (UV and visible)
2. Wavelength selector (monochromator)
3. Sample containers
4. Detector
5. Signal processor and readout
Instrumental components
Sources of UV radiation
It is important that the power of the radiation source does not change abruptly over it's
wavelength range.
The electrical excitation of deuterium or hydrogen at low pressure produces a continuous UV
spectrum. The mechanism for this involves formation of an excited molecular species, which
breaks up to give two atomic species and an ultraviolet photon. This can be shown as;
D2 + electrical energy D2 * D' + D'' + hv
Both deuterium and hydrogen lamps emit radiation in the range 160 - 375 nm. Quartz
windows must be used in these lamps, and quartz cuvettes must be used, because glass
absorbs radiation of wavelengths less than 350 nm.
Introduction
Sources of visible radiation
The tungsten filament lamp is commonly employed as a source of visible light. This type of
lamp is used in the wavelength range of 350 - 2500 nm. The energy emitted by a tungsten
filament lamp is proportional to the fourth power of the operating voltage. This means that
for the energy output to be stable, the voltage to the lamp must be very stable indeed.
Electronic voltage regulators or constant-voltage transformers are used to ensure this
Tungsten/halogen lamps contain a small amount of iodine in a quartz "envelope" which also
contains the tungsten filament. The iodine reacts with gaseous tungsten, formed by
sublimation, producing the volatile compound WI 2. When molecules of WI2 hit the filament
they decompose, redepositing tungsten back on the filament. The lifetime of a
tungsten/halogen lamp is approximately double that of an ordinary tungsten filament lamp.
Tungsten/halogen lamps are very efficient, and their output extends well into the ultra-violet.
They are used in many modern spectrophotometers.
Wavelength selector (monochromator)
All monochromators contain the following component parts;
An entrance slit
A collimating lens
A dispersing device (usually a prism or a grating)
A focusing lens
An exit slit
Polychromatic radiation (radiation of more than one wavelength) enters the monochromator
through the entrance slit. The beam is collimated, and then strikes the dispersing element at
an angle. The beam is split into its component wavelengths by the grating or prism. By
moving the dispersing element or the exit slit, radiation of only a particular wavelength
leaves the monochromator through the exit slit.
Czerney-Turner grating monochromator
stability.
Cuvettes
The containers for the sample and reference solution must be transparent to the radiation
which will pass through them. Quartz or fused silica cuvettes are required for spectroscopy in
the UV region. These cells are also transparent in the visible region. Silicate glasses can be
used for the manufacture of cuvettes for use between 350 and 2000 nm.
Detectors
The photomultiplier tube is a commonly used detector in UV-Vis spectroscopy. It consists
of a photoemissive cathode (a cathode which emits electrons when struck by photons of
radiation), several dynodes (which emit several electrons for each electron striking them) and
an anode.
A photon of radiation entering the tube strikes the cathode, causing the emission of several
electrons. These electrons are accelerated towards the first dynode (which is 90V more
positive than the cathode). The electrons strike the first dynode, causing the emission of
several electrons for each incident electron. These electrons are then accelerated towards the
second dynode, to produce more electrons which are accelerated towards dynode three and so
on. Eventually, the electrons are collected at the anode. By this time, each original photon has
produced 106 - 107 electrons. The resulting current is amplified and measured.
Photomultipliers are very sensitive to UV and visible radiation. They have fast response
times. Intense light damages photomultipliers; they are limited to measuring low power
radiation.
Cross section of a photomultiplier tube
The linear photodiode array is an example of a multichannel photon detector. These

detectors are capable of measuring all elements of a beam of dispersed radiation
simultaneously.
A linear photodiode array comprises many small silicon photodiodes formed on a single
silicon chip. There can be between 64 to 4096 sensor elements on a chip, the most common
being 1024 photodiodes. For each diode, there is also a storage capacitor and a switch. The
individual diode-capacitor circuits can be sequentially scanned.
In use, the photodiode array is positioned at the focal plane of the monochromator (after the
dispersing element) such that the spectrum falls on the diode array. They are useful for
recording UV-Vis. absorption spectra of samples that are rapidly passing through a sample
flow cell, such as in an HPLC detector.
Charge-Coupled Devices (CCDs) are similar to diode array detectors, but instead of diodes,
they consist of an array of photocapacitors.
2. Explain the construction and working of UV-Visible Spectrophotometer
Introduction
If you pass white light through a coloured substance, some of the light gets
absorbed. A solution containing hydrated copper(II) ions, for example, looks pale
remaining wavelengths in the light combine in the eye and brain to give the
appearance of cyan (pale blue).
Some colourless substances also absorb light - but in the ultra-violet region. Since
we can't see UV light, we don't notice this absorption.
Different substances absorb different wavelengths of light, and this can be used to
help to identify the substance - the presence of particular metal ions, for example, or
of particular functional groups in organic compounds.
The amount of absorption is also dependent on the concentration of the substance if
it is in solution. Measurement of the amount of absorption can be used to find
concentrations of very dilute solutions.
An absorption spectrometer measures the way that the light absorbed by a
compound varies across the UV and visible spectrum.
A simple double beam spectrometer
We'll start with the full diagram, and then explain exactly what is going on at each
stage.
blue because the solution absorbs light from the red end of the spectrum. The
Working
The light source
You need a light source which gives the entire visible spectrum plus the near ultraviolet so that you are covering the range from about 200 nm to about 800 nm. (This
extends slightly into the near infra-red as well.)
You can't get this range of wavelengths from a single lamp, and so a combination of
two is used - a deuterium lamp for the UV part of the spectrum, and a tungsten /
halogen lamp for the visible part.
The combined output of these two bulbs is focussed on to a diffraction grating.
The diffraction grating and the slit

You are probably familiar with the way that a prism splits light into its component
colours. A diffraction grating does the same job, but more efficiently.
The blue arrows show the way the various wavelengths of the light are sent off in
wavelengths through into the rest of the spectrometer.

By gradually rotating the diffraction grating, you can allow light from the whole
spectrum (a tiny part of the range at a time) through into the rest of the instrument.
The rotating discs

This is the clever bit! Each disc is made up of a number of different segments.
Those in the machine we are describing have three different sections - other designs
may have a different number.
The light coming from the diffraction grating and slit will hit the rotating disc and
one of three things can happen.
1. If it hits the transparent section, it will go straight through and pass through
the cell containing the sample. It is then bounced by a mirror onto a second
rotating disc.
This disc is rotating such that when the light arrives from the first disc, it
different directions. The slit only allows light of a very narrow range of
meets the mirrored section of the second disc. That bounces it onto the
detector.
2. If the original beam of light from the slit hits the mirrored section of the first
rotating disc, it is bounced down along the green path. After the mirror, it
passes through a reference cell (more about that later).
Finally the light gets to the second disc which is rotating in such a way that
it meets the transparent section. It goes straight through to the detector.
3. If the light meets the first disc at the black section, it is blocked - and for a
very short while no light passes through the spectrometer. This just allows
the computer to make allowance for any current generated by the detector in
the absence of any light.
It is following the red path in the diagram:
The sample and reference cells
These are small rectangular glass or quartz containers. They are often designed so
that the light beam travels a distance of 1 cm through the contents.
The sample cell contains a solution of the substance you are testing - usually very
in the wavelength range we are interested in (200 - 800 nm).

The reference cell just contains the pure solvent.
The detector and computer

The detector converts the incoming light into a current. The higher the current, the
greater the intensity of the light.
For each wavelength of light passing through the spectrometer, the intensity of the
light passing through the reference cell is measured. This is usually referred to as
Io - that's I for Intensity.
The intensity of the light passing through the sample cell is also measured for that
wavelength - given the symbol, I.
If I is less than Io, then obviously the sample has absorbed some of the light. A
simple bit of maths is then done in the computer to convert this into something
called the absorbance of the sample - given the symbol, A.
An absorbance of 0 at some wavelength means that no light of that particular

wavelength has been absorbed. The intensities of the sample and reference beam are
both the same, so the ratio Io/I is 1. Log10 of 1 is zero.
dilute. The solvent is chosen so that it doesn't absorb any significant amount of light
An absorbance of 1 happens when 90% of the light at that wavelength has been
absorbed - which means that the intensity is 10% of what it would otherwise be.
In that case, Io/I is 100/I0 (=10) and log10 of 10 is 1.
The chart recorder
like this:
This particular substance has what are known as absorbance peaks at 255 and 395
nm.
3.
Explain the construction and working of FTIR?
Introduction
FTIR spectrometers (Fourier Transform Infrared Spectrometer) are widely used in organic
synthesis, polymer science, petrochemical engineering, pharmaceutical industry and food
analysis. In addition, since FTIR spectrometers can be hyphenated to chromatography, the
mechanism of chemical reactions and the detection of unstable substances can be investigated
with such instruments. FTIR spectrometers (Fourier Transform Infrared Spectrometer) are
widely
used
in
organic
synthesis,
polymer
science,
petrochemical
engineering,
pharmaceutical industry and food analysis. In addition, since FTIR spectrometers can be
Chart recorders usually plot absorbance against wavelength. The output might look
hyphenated to chromatography, the mechanism of chemical reactions and the detection of
unstable substances can be investigated with such instruments.
The Components of FTIR Spectrometers
A common FTIR spectrometer consists of a source, interferometer, sample compartment,
detector, amplifier, A/D convertor, and a computer. The source generates radiation which
amplified and converted to digital signal by the amplifier and analog-to-digital converter,
respectively. Eventually, the signal is transferred to a computer in which Fourier transform is
carried out. Figure 2 is a block diagram of an FTIR spectrometer.
Figure 2. Block diagram of an FTIR spectrometer

Michelson Interferometer
The Michelson interferometer, which is the core of FTIR spectrometers, is used to split one
beam of light into two so that the paths of the two beams are different. Then the Michelson
interferometer recombines the two beams and conducts them into the detector where the
difference of the intensity of these two beams are measured as a function of the difference of
the paths. Figure 3 is a schematic of the Michelson Interferometer.
passes the sample through the interferometer and reaches the detector. Then the signal is
Figure 3. Schematic of the Michelson interferometer
A typical Michelson interferometer consists of two perpendicular mirrors and a beamsplitter.

One of the mirror is a stationary mirror and another one is a movable mirror. The
beamsplitter is designed to transmit half of the light and reflect half of the light.
Subsequently, the transmitted light and the reflected light strike the stationary mirror and the
movable mirror, respectively. When reflected back by the mirrors, two beams of light
recombine with each other at the beamsplitter.
If the distances travelled by two beams are the same which means the distances between two
mirrors and the beamsplitter are the same, the situation is defined as zero path difference
(ZPD). But imagine if the movable mirror moves away from the beamsplitter, the light beam
which strikes the movable mirror will travel a longer distance than the light beam which
strikes the stationary mirror. The distance which the movable mirror is away from the ZPD is
defined as the mirror displacement and is represented by . It is obvious that the extra
distance travelled by the light which strikes the movable mirror is 2. The extra distance is
defined as the optical path difference (OPD) and is represented by delta. Therefore,
=2
It is well established that when OPD is the multiples of the wavelength, constructive
interference occurs because crests overlap with crests, troughs with troughs. As a result, a
maximum intensity signal is observed by the detector. This situation can be described by the
following equation:
(n = 0,1,2,3...)
In contrast, when OPD is the half wavelength or half wavelength add multiples of
wavelength, destructive interference occurs because crests overlap with troughs.
Consequently, a minimum intensity signal is observed by the detector. This situation can be
described by the following equation:
=(n+12)
(n = 0,1,2,3...)
These two situations are two extreme situations. If the OPD is neither n-fold wavelengths nor
(n+1/2)-fold wavelengths, the interference should be between constructive and destructive.
So the intensity of the signal should be between maximum and minimum. Since the mirror
moves back and forth, the intensity of the signal increases and decreases which gives rise to a
cosine wave. The plot is defined as an interferogram. When detecting the radiation of a broad
band source rather than a single-wavelength source, a peak at ZPD is found in the
interferogram. At the other distance scanned, the signal decays quickly since the mirror
moves back and forth. Figure 4(a)shows an interferogram of a broad band source.
Fourier Transform of Interferogram to Spectrum

The interferogram is a function of time and the values outputted by this function of time are
said to make up the time domain. The time domain is Fourier transformed to get a frequency
domain, which is deconvoluted to product a spectrum. Figure 4 shows the Fast Fourier
transform from an interferogram of polychromatic light to its spectrum.
=n
(a)
(b)
Figure 4. (a) Interferogram of a monochromatic light; (b) its spectrum
4.
Derive Beer Lamberts law and explain about deviations from Beers Law?
Introduction
The Beer-Lambert law (also called the Beer-Lambert-Bouguer law or simply Beer's law) is
the linear relationship between absorbance and concentration of an absorber of
electromagnetic radiation. The general Beer-Lambert law is usually written as:
where A is the measured absorbance, a
is a wavelength-dependent absorptivity coefficient,
b is the path length, and c is the analyte concentration. When working in concentration units
of molarity, the Beer-Lambert law is written as:
where
The
is the wavelength-dependent molar absorptivity coefficient with units of M -1 cm-1.

subscript is often dropped with the understanding that a value for
is for a specific
wavelength. If multiple species that absorb light at a given wavelength are present in a
sample, the total absorbance at that wavelength is the sum due to all absorbers:
where the subscripts refer to the molar absorptivity and concentration of the different
absorbing species that are present.
Theory
as:
where P is the power of light after it passes through the sample and P o is the initial light
power. The relation between A and T is:
The figure shows the case of absorption of light through an optical filter and includes other
processes that decreases the transmittance such as surface reflectance and scattering.
Experimental measurements are usually made in terms of transmittance (T), which is defined
Derivation of the Beer-Lambert law
The Beer-Lambert law can be derived from an approximation for the absorption coefficient
for a molecule by approximating the molecule by an opaque disk whose cross-sectional area,
, represents the effective area seen by a photon of frequency w. If the frequency of the light
is far from resonance, the area is approximately 0, and if w is close to resonance the area is a
Io is the intensity entering the sample at z=0, I z is the intensity entering the infinitesimal slab
at z, dI is the intensity absorbed in the slab, and I is the intensity of light leaving the sample.
Then, the total opaque area on the slab due to the absorbers is
fraction of photons absorbed will be
* N * A * dz. Then, the
* N * A * dz / A so,
Integrating this equation from z = 0 to z = b gives:
maximum. Taking an infinitesimal slab, dz, of sample:
or
Since N (molecules/cm3) * (1 mole / 6.023x1023 molecules) * 1000 cm3 / liter = c
or
where
* (6.023x1020 / 2.303) =
* 2.61x1020
Limitations of the Beer-Lambert law

The linearity of the Beer-Lambert law is limited by chemical and instrumental factors. Causes
of nonlinearity include:
deviations in absorptivity coefficients at high concentrations (>0.01M) due to
electrostatic interactions between molecules in close proximity
scattering of light due to particulates in the sample
fluoresecence or phosphorescence of the sample
changes in refractive index at high analyte concentration
shifts in chemical equilibria as a function of concentration
non-monochromatic radiation, deviations can be minimized by using a relatively flat
part of the absorption spectrum such as the maximum of an absorption band
stray light
(moles/liter) and 2.303 * log(x) = ln(x), then
5. Explain with neat diagram the construction and working of atomic absorption
spectrophotometer?
A diagram of a Flame Atomic Absorption Spectrometer is shown in figure 11. The light
source is a cold cathode lamp that produces (almost exclusively) the light that would be
naturally emitted by the element to be measured at a high temperature. A large range of such
lamps are available that includes the vast majority of the elements of general analytical
in the flame will selectively absorb. The light passes through the flame, which is usually
rectangular in shape so as to provide an adequate path length of flame for the light to be
absorbed, and then into the optical system of the spectrometer.
The flame is fed with a combustible gas, customarily air/acetylene, nitrous oxide/acetylene or
air/propane or butane. The sample, dissolved in a suitable solvent, is nebulized and fed into
the gas stream at the base of the burner. The light, having passed through the flame, can be
focused directly onto a photo-cell or onto a Diffraction Grating by means of a spherical
mirror. The Diffraction Grating can be made movable, and so it can be set to monitor a
particular wavelength that is characteristic of the element being measured, or it can be
scanned to produce a complete absorption Spectrum of the sample. After leaving the grating,
light of a selected wavelength, or range of wavelengths, is focused onto the photocell. The
position of the Diffraction Grating determines the wavelength of the light that is to be
monitored.
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interest. Consequently, the light will contain specifically those wavelengths that the element
Figure 1. The Basic System of a Flame Atomic Absorption Spectrometer

The flame absorption spectrometer is fairly sensitive, and can be readily used as a tandem
instrument combined with a chromatograph, providing that an appropriate interface is
employed. This instrument is normally fitted with 4 different cold cathode lamps and, thus,
can examine 4 different elements, automatically, from the same analysis; 8 lamp units are
also available. A diagram of a hollow cathode lamp is shown in figure 13.
The cylindrical hollow cathode of the lamp contains one or more of the elements of interest in
the analysis. The cylindrical cathode is screened from the anode connections by means of a
ceramic cylinder The anode is situated above the cathode and is made of tungsten or nickel
and the whole electrode system is enclosed in a glass envelope. The glass envelope is filled
with neon or argon to a pressure of 1kPa and on applying a potential of 100 to 200 volts
across the electrodes a glow discharge is formed.
Figure 2. The Hollow Cathode Element Lamp.

A simple explanation of the process is as follows, Accelerated electrons from the cathode
collide with the gas atoms and produce ions. The ions are accelerated to the cathode by the
electric field and when they strike the cathode surface the elements of interest are ejected
from the surface and are excited to provide radiation in the discharge environment. These
lamps can be combined in groups so that a given instrument can determine a number of
different elements by merely switching the lamps.
A single lamp can be made to generate characteristic radiation for up to two or three elements
without interference problems. Modern atomic adsorption spectrometers are deigned to
provide extremely simple operation, easy maintenance and, when required, fast and
inexpensive servicing. Many ancillary and alternative devices are available to suit specific
applications.
A number of different sampling devices are available including very sophisticated automatic
samplers that allow instruments to be operated 24 hours a day.
With a neat sketch explain flame emission spectroscopy
The analytical apparatus

The functions of an analytical flame spectrometer in general are:
(a) transformation of the solution to be analyzed into a vapour containing free atoms
or molecular compounds of the analyte in the flame;
(b) Selection and detection of the optical signal (arising from the analyte vapour)
which carries information on the kind and concentration of the analyte;
(c) Amplification and read-out of the electrical signal.
Transformation of sample into vapour
With a pneumatic nebulizer operated by a compressed gas, the solution is aspirated
from the sample container and nebulized into a mist or aerosol of fine droplets.
By
desolvation, i.e., evaporation of the solvent from the droplets, this mist is
converted into a dry aerosol which is volatilized in the flame.

The atomization, i.e., the conversion ofvolatilized analyte into free atoms is
performed by the flame or other atomizer.
The total consumption time is the time required to consume the sample entirely. The
minimum consumption time is the time for which nebulization must be carried out to
perform an analysis with a given precision.
Nebulizers can be described as follows:
According to the source of energy used for nebulization as, for example,
6.
pneumatic or ultrasonic nebulizers.
According to the way the liquid is taken up, e.g., suction, gravity-fed, controlled
flow, and reflux-nebulizers.
According to the relative position of the capillaries for the nebulizing gas and the
aspirated liquid, e.g., angular and concentric nebulizers.
In the chamber-type nebulizer, the nebulizing gas-jet stream emerges from the sprayer
Special devices are the nebulizer with heated spray chamber, the
twin nebulizer, and the drop generator.
Construction and working:

Flames are produced by means of a burner to which fuel and oxidant are supplied in
thenform of gases.
With the premix burner, fuel and oxidant are thoroughly mixed inside the burner
housing before they leave the burner ports and enter the primary combustion or inner
zone of the flame. This type of burner usually produces an approximately laminar
flame, and is commonly combined with a separate unit for nebulizing the sample.
In contrast, a direct-injection burner combines the function of nebulizer and burner.
Here oxidant and fuel emerge from separate ports and are mixed above the burner
orifice to produce a turbulent flame. Most commonly, the oxidant is also used for
aspirating and nebulizing the sample.
However, when the fuel is used for this purpose, the term reversed direct-injection
burner is applied. In each case, the mist droplets enter the flame directly, without
passing through a spray chamber.
The term total-consumption burner, which is often used, is not recommended.
Premix burners are distinguished as Bunsen-, Meker-, or slot-burners according to
whether they have one large hole, a number of small holes, or a slot as outlet for the
gas mixture, respectively. When several parallel slots are present, they are identified
as multislot burners (e.g., a three-slot burner).
The small diameter of the holes in the Meker burner or the narrowness of the slot in
the slot-burner prevents the unwanted flash-back of the flame into the burner housing.
At the edge of the flame where the hot gas comes into contact with the surrounding
air, secondary combustion occurs and the secondary combustion or outer zone is
formed.
into a spray chamber.
The region of the flame confined by the inner and outer zones, where in many
instances the conditions for flame analysis are optimum, is called the interzonal
region, or, when the combustion zones have the form of a cone, the interconal zone.
Sometimes provision is made to screen the observed portion of the flame gases from
direct contact with the surrounding air. This may be done either mechanically, by
placing a tube on the top of the burner around the flame, which produces a zonal
sheath of inert gas that emerges from openings at the rim of the burner top (shielded
flame).
Observations can thus be made without disturbances from the secondary-combustion
zone.
separation (separated flame), or aerodynamically, by surrounding the flame with a
Unit II
CHROMATOGRAPHY
1. What is chromatogaraphy ?
chromatography is a physical method of seperation of the commponents of a mixture
by distribution between two phases, of which one is stationary bed of a large surface
area and other fluid phase (Mobile phase) that percolates through or along the
stationay bed.
2. What are the types of chromatography?
Chromatography
Gas
chromatography
Gas-Liquid (GLC)
Gas_Solid (GSC)
Liquid
Chromatography
Liquid-Liquid
(LLC)
Bonded Phase
(BPC)
Liqid _Solid (LSC)
Ion Exchange
(IEC)
Exclusioin (EC)
Ion Pair
Chromatography
3. What is Retention Time?

The retention time is the total time that a compound spends in both the mobile phase
and stationary phase.
The time between sample injecction and an analyte peak reaching a detector at the end
of the column is termed as the retention time (t R).
4. What is Gas Chromatography ?
Gas Chrromatography is an analytical technique used for seperating compounds based
primarily on their volatalities. Gas chromatography provides both qualitative and
quantitative information for individual compounds present in a sample.
5. List out the types of colummns used in gas chromatography
Packed Column
Capillary Column
6. List out the various detectors used in gas chromatography
The katharometer or Thermal conductivity detector
Flame ionization Detectors
PART A
Flame Photometric Detectors

7. What are the advantages of Gas Chromatography?
High flow rates of mobile phase is possible
Several methods of detecting components in flowing gas stream are available
8. What is meant by efficiency in chromatography?
The efficiency is related to the number of compounds that can be seperated by the
column.It is expressed as the number of Theoritical plates or as the highest equivalent
to a theortical plate.
9. Why High pressure pumps are used in HPLC?
Liquid chromatography consists of columns with packing, through which the mobile
phase and the sample flows. The packing in the column reduces the flow rate of the
mobile phase. In order to ensure constant flow rate high pressure pumps are used to
pump the mobile phase into the column.
10. What are the advantages of Katharometer ?
It is a non destructive method of testing
High sensitivity can be achieved, which enables us to detect small quanitiy of
a compound coming out of a mixture
The measurement errors are minimized
PART B
1. With a neat block diagram explain the construction and working of gas
chromatography
Introduction
Gas chromatography - specifically gas-liquid chromatography - involves a sample being
vapourised and injected onto the head of the chromatographic column. The sample is
transported through the column by the flow of inert, gaseous mobile phase. The column itself
contains a liquid stationary phase which is adsorbed onto the surface of an inert solid.
Instrumental components
Carrier gas
The carrier gas must be chemically inert. Commonly used gases include nitrogen, helium,
argon, and carbon dioxide. The choice of carrier gas is often dependant upon the type of
detector which is used. The carrier gas system also contains a molecular sieve to remove
Sample injection port

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. For packed columns, sample
size ranges from tenths of a microliter up to 20 microliters. Capillary columns, on the other
hand, need much less sample, typically around 10 -3 mL. For capillary GC, split/splitless
injection is used. Have a look at this diagram of a split/splitless injector;
water and other impurities.
The injector can be used in one of two modes; split or splitless. The injector contains a heated
chamber containing a glass liner into which the sample is injected through the septum. The
carrier gas enters the chamber and can leave by three routes (when the injector is in split
mode). The sample vapourises to form a mixture of carrier gas, vapourised solvent and
vapourised solutes. A proportion of this mixture passes onto the column, but most exits
through the split outlet. The septum purge outlet prevents septum bleed components from
Columns
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 1.5 - 10m
in length and have an internal diameter of 2 - 4mm.
Capillary columns have an internal diameter of a few tenths of a millimeter. They can be one
of two types; wall-coated open tubular (WCOT) or support-coated open tubular (SCOT).
Wall-coated columns consist of a capillary tube whose walls are coated with liquid stationary
phase. In support-coated columns, the inner wall of the capillary is lined with a thin layer of
support material such as diatomaceous earth, onto which the stationary phase has been
adsorbed. SCOT columns are generally less efficient than WCOT columns. Both types of
capillary column are more efficient than packed columns.
In 1979, a new type of WCOT column was devised - the Fused Silica Open Tubular (FSOT)
column;
entering the column.
These have much thinner walls than the glass capillary columns, and are given strength by the
polyimide coating. These columns are flexible and can be wound into coils. They have the
advantages of physical strength, flexibility and low reactivity.
For precise work, column temperature must be controlled to within tenths of a degree. The
optimum column temperature is dependant upon the boiling point of the sample. As a rule of
thumb, a temperature slightly above the average boiling point of the sample results in an
elution time of 2 - 30 minutes. Minimal temperatures give good resolution, but increase
elution times. If a sample has a wide boiling range, then temperature programming can be
useful. The column temperature is increased (either continuously or in steps) as separation
proceeds.
Detectors
There are many detectors which can be used in gas chromatography. Different detectors will
give different types of selectivity. A non-selective detector responds to all compounds except
the carrier gas, a selective detector responds to a range of compounds with a common
physical or chemical property and a specific detector responds to a single chemical
compound. Detectors can also be grouped into concentration dependant detectors and mass
flow dependant detectors. The signal from a concentration dependant detector is related to the
concentration of solute in the detector, and does not usually destroy the sample Dilution of
with make-up gas will lower the detectors response. Mass flow dependant detectors usually
destroy the sample, and the signal is related to the rate at which solute molecules enter the
detector. The response of a mass flow dependant detector is unaffected by make-up gas.
Column temperature
The effluent from the column is mixed with hydrogen and air, and ignited. Organic
compounds burning in the flame produce ions and electrons which can conduct electricity
through the flame. A large electrical potential is applied at the burner tip, and a collector
electrode is located above the flame. The current resulting from the pyrolysis of any organic
compounds is measured. FIDs are mass sensitive rather than concentration sensitive; this
gives the advantage that changes in mobile phase flow rate do not affect the detector's
response. The FID is a useful general detector for the analysis of organic compounds; it has
high sensitivity, a large linear response range, and low noise. It is also robust and easy to use,
but unfortunately, it destroys the sample.
2. With a help of neat sketch, explain the construction and working of Katharometer
The thermal conductivity detector (TCD), also known as a Katharometer, is a bulk
property detector and a chemical specific detector commonly used in gas chromatography.
[1]
This detector senses changes in the thermal conductivity of the column effluent and
compares it to a reference flow of carrier gas. Since most compounds have a thermal
conductivity much less than that of the common carrier gases of helium or hydrogen, when an
analyte elutes from the column the effluent thermal conductivity is reduced, and a detectable
signal is produced.
Operation
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. When an
analyte elutes and the thermal conductivity of the column effluent is reduced, the filament
heats up and changes resistance. This resistance change is often sensed by a Wheatstone
bridge circuit which produces a measurable voltage change. The column effluent flows over
circuit.
TCD Schematic
A schematic of a classic thermal conductivity detector design utilizing a wheatstone bridge
circuit is shown. The reference flow across resistor 4 of the circuit compensates for drift due
to flow or temperature fluctuations. Changes in the thermal conductivity of the column
effluent flow across resistor 3 will result in a temperature change of the resistor and therefore
a resistance change which can be measured as a signal.
Since all compounds, organic and inorganic, have a thermal conductivity different from
helium, all compounds can be detected by this detector. The TCD is often called a universal
detector because it responds to all compounds. Also, since the thermal conductivity of
organic compounds are similar and very different from helium, a TCD will respond similarly
to similar concentrations of analyte. Therefore the TCD can be used without calibration and
one of the resistors while the reference flow is over a second resistor in the four-resistor
the concentration of a sample component can be estimated by the ratio of the analyte peak
area to all components (peaks) in the sample.
The TCD is a good general purpose detector for initial investigations with an unknown
sample. Since the TCD is less sensitive than the flame ionization detector and has a larger
dead volume it will not provide as good resolution as the FID. However, in combination with
thick film columns and correspondingly larger sample volumes, the overall detection limit
specific and non-destructive.

The TCD is also used in the analysis of permanent gases (argon, oxygen, nitrogen, carbon
dioxide) because it responds to all these pure substances unlike the FID which cannot detect
compounds which do not contain carbon-hydrogen bonds.
Process description
It functions by having two parallel tubes both containing gas and heating coils. The gases are
examined by comparing the rate of loss of heat from the heating coils into the gas. The coils
are arranged in a bridge circuit so that resistance changes due to unequal cooling can be
measured. One channel normally holds a reference gas and the mixture to be tested is passed
through the other channel.
Applications
In the oil industry katharometers have been used for a long time for hydrocarbon detection
but have a history of unstable calibrations in non-stationary oil-related applications. In
normal drilling practice, 5 hydrocarbon gases, plus a couple of non-hydrocarbon gases, are
expected in normal samples resulting in cross-talk between the methane absorption line and
the ethane. Hence the current use of flame ionization detectors.
Katharometers are used medically in lung function testing equipment and in gas
chromatography. The results are slower to obtain compared to a mass spectrometer, but the
device is inexpensive, and has good accuracy when the gases in question are known, and it is
only the proportion that must be determined.
Monitoring of hydrogen purity in hydrogen-cooled turbogenerators
can be similar to that of an FID. The TCD is not as sensitive as other detectors but it is non-
3. Explain with a neat diagram the construction and working of flame ionization
detector
A flame ionization detector (FID) is a scientific instrument that measures the concentration
of organic species in a gas stream. It is frequently used as a detector in gas chromatography.
Standalone FIDs can also be used in applications such as landfill gas monitoring and fugitive
Operating principle
The operation of the FID is based on the detection of ions formed during combustion of
organic compounds in a hydrogen flame. The generation of these ions is proportional to the
concentration of organic species in the sample gas stream. Hydrocarbons generally have
molar response factors that are equal to number of carbon atoms in their molecule, while
oxygenates and other species that contain heteroatoms tend to have a lower response factor.
Carbon monoxide and carbon dioxide are not detectable by FID.
emissions monitoring in stationary or portable instruments.
FID Schematic
The design of the flame ionization detector varies from manufacturer to manufacturer, but the
principles are the same. Most commonly, the FID is attached to a gas chromatography
system.
The eluent exits the GC column (A) and enters the FID detectors oven (B). The oven is
needed to make sure that as soon as the eluent exits the column, it does not come out of the
gaseous phase and deposit on the interface between the column and FID. This deposition
would result in loss of eluent and errors in detection. As the eluent travels up the FID, it is
first mixed with the hydrogen fuel (C) and then with the oxidant (D). The eluent/fuel/oxidant
mixture continues to travel up to the nozzle head where a positive bias voltage exists (E).
This positive bias helps to repel the reduced carbon ions created by the flame (F) pyrolyzing
the eluent. The ions are repelled up toward the collector plates (G) which are connected to a
very sensitive ammeter, which detects the ions hitting the plates, then feeds that signal (H) to
Description of a generic detector
an amplifier, integrator, and display system. The products of the flame are finally vented out
of the detector through the exhaust port (J).
Operation
In order to detect these ions, two electrodes are used to provide a potential difference. The
positive electrode doubles as the nozzle head where the flame is produced. The other,
was either tear-drop shaped or angular piece of platinum. Today, the design has been
modified into a tubular electrode, commonly referred to as a collector plate. The ions thus are
attracted to the collector plate and upon hitting the plate, induce a current. This current is
measured with a high-impedance picoammeter and fed into an integrator. The manner in
which the final data is displayed is based on the computer and software. In general, a graph is
displayed that has time on the x-axis and total ion on the y-axis.
The current measured corresponds roughly to the proportion of reduced carbon atoms in the
flame. Specifically how the ions are produced is not necessarily understood, but the response
of the detector is determined by the number of carbon atoms (ions) hitting the detector per
unit time. This makes the detector sensitive to the mass rather than the concentration, which
is useful because the response of the detector is not greatly affected by changes in the carrier
gas flow rate.
Advantages
Flame ionization detectors are used very widely in gas chromatography because of a number
of advantages.
Cost: Flame ionization detectors are relatively inexpensive to acquire and operate.
Low maintenance requirements: Apart from cleaning or replacing the FID jet, these
detectors require no maintenance.
Rugged construction: FIDs are relatively resistant to misuse.
Linearity and detection ranges: FIDs can measure organic substance concentration at
very low and very high levels, having a linear response of 10^6.
negative electrode is positioned above the flame. When first designed, the negative electrode
Disadvantages
Flame ionization detectors cannot differentiate between different organic substances. They
also cannot detect inorganic substances. In some systems, CO and CO2 can be detected in the
FID using a methanizer, which is a bed of Ni catalyst that reduces CO and CO2 to methane,
which can be in turn detected by the FID.
through it; all hydrocarbons and oxygenates are oxidized to carbon dioxide and water and
other heteroatoms are oxidized according to thermodynamics. For this reason, FIDs tend to
be the last in a detector train and also cannot be used for preparatory work.
4. Explain the construction and working of Ion exchange chromatography with a neat
diagram
Ion exchange means exchange of ions from a medium. It has typical application in water
softening by exchange of alkaline metal ions like Ca2+, Mg2+ by Na+. Other common
applications are sugar processing, hydrometallurgical application, protein fractionation,
biological separation, etc.
Fundamentals:
Ion from a solution is removed when it is passed through a bed of exchangeable ions, called
resins.
In this reaction, R- is fixed negative charge on the resin. A+ and B+ are called counter-ions
and X- is called coion in resin phase
Ion Exchange Resin
Most popular base for ion exchange resin is polystyrene. Cross linking with divinylbenzene
(DVB) is done with resin to make it insoluble. About 2-10% DVB is used. Both (i)
macroporous
and
(ii)
gel
type
resin
beads
are
used.
Macroporous beads have pores inside the beads where ions can go in or get out.
Another important disadvantage is that the FID flame oxidizes all compounds that pass
Typical external porosity is about 0.40. Gel type resin have various degrees of swelling.
These may be polystyrene-sulfuric acid resin with various % of DVB, polyacrylic acid resin,
etc.
Acidic resins have negative fixed charges and can exchange cations. Basic resins have
positive fixed charges and can exchange anions. Exchangers can also weak or strong. Strong
resins are fully ionized and all the fixed groups are available to exchange cations. Strong base
can
degrade
at
higher
pH
and
temperature.
On the other hand, weak resins are only partially ionized at most pH values. They
have lower exchange capacity but they are easier to regenerate. Weak resins require less
regenerant than strong resins.
But weak resins swell or contract when ions are exchanged. They can rupture due to improper
stress distribution of during expansion/contraction cycle. In weak resin also the ions diffuse
slowly. So, mass transfer resistance is very high and time requirement is long.
Techniques of Ion Exchange Chromatography:
1. Preparation of Column
The ion exchange chromatography is carried out in a chromatographic column which usually
consists of a burette provided with a glass wool plug at the lower end. Generally a ratio of 10:
1 or 100:1 between height and diameter is maintained in most of the experiment. Too narrow
or too wide column give uneven flow of liquid and sometimes poor separation.
2. Preparation of Ion Exchange

Ion exchange materials are first allowed to swell in buffer or in HCl or NaOH solution for 2-3
hours or sometimes overnight. Almost all ion exchange resin swells when placed in buffer or
distilled water and this is due to hydration of their ions. In dry condition, the pore of resins is
restricted so in order to swell the pore of resin. Resins are suspended in buffer solution or in
distilled water.
3. Washing of Ion Exchangers
resins
The ion exchange material is obtained in required ionic form by washing with appropriate
solution. For e.g. the H+ form of cation exchange resins is obtained by washing the material
with HCl then with water until the washings are neutral.
Anionic exchangers are generally supplied in the form of salt and amines. Similarly, Na +
Figure 2: Ion exchange chromatography

4. Packing of Column
This is one of the most critical factors in achieving a successful separation. The column is
held in vertical position and the slurry of resins is poured into the column that has its outlet
closed. The column is gently tapped to ensure that no air bubbles are trapped and that packing
material settles evenly.
5. Sample Application
Sample can be loaded by using pipette or syringe. The amount of sample that can be applied
to a column is dependent upon the size of the column and the capacity of resins, If the
starting buffer is to be used throughout the development of column, the sample volume be 1
% to 5 % of bed volume.
6. Development an Elution of bound ions
form is prepared by washing the resins with NaCl or NaOH solution and then with water.
Bound ions can be removed by changing the pH of buffer. E.g. separation of amino acid is
usually achieved by using a strong acidic cation exchanger. The sample is introduced onto the
column at pH of 1-2, thus ensuring complete binding of all of the amino acids.
Gradient elution used in increasing pH and ionic concentration results in the sequential
elution of amino acid. Then acidic amino acid such as aspartic acid and glutamic acid are
eluted first. The neutral amino acid such as glycine and valine are eluted. The basic amino
eluted at last.
7. Analysis of eluate
Equal fraction of each elute are collected at different test tube keeping the flow rate at 1 ml
per minute. The eluate collected in each fraction is mixed with ninhydrin color reagent. The
mixture is then heated to 105 C to develop the color and intensity of color is determined by
colorimeter method or spectrophotometer method at 540 to 570 nm.
5.Explain the construction and working of High Pressure Liquid Chromatography

(HPLC) with a neat sketch
Introduction
High-performance liquid chromatography (HPLC) is a form of liquid chromatography to
separate compounds that are dissolved in solution. HPLC instruments consist of a reservoir of
mobile phase, a pump, an injector, a separation column, and a detector. Compounds are
separated by injecting a plug of the sample mixture onto the column. The different
components in the mixture pass through the column at different rates due to differences in
their partitioning behavior between the mobile liquid phase and the stationary phase.
Instrumentation
Solvents must be degassed to eliminate formation of bubbles. The pumps provide a steady
high pressure with no pulsating, and can be programmed to vary the composition of the
solvent during the course of the separation.
The liquid sample is introduced into a sample loop of an injector with a syringe. When the
loop is filled, the injector can be inject the sample into the stream by placing the sample loop
in line with the mobile phase tubing.
acid such as lysine and arginine retain their net positive charge at pH value of 9 to 11 and are
The presence of analytes in the column effluent is recorded by detecting a change in
refractive index, UV-VIS absorption at a set wavelength, fluorescence after excitation with a
suitable wavelength, or electrochemical response.
Mass spectrometers can also be interfaced with liquid chromatography to provide structural
information and help identify the separated analytes.
Advantages
Speed (minutes)
High resolution
Sensitivity (ng to fg)
Reproducibility of +/- 1%
Accuracy
Automation
Schematic of an HPLC instrument
Disadvantages
Cost
Complexity
Low sensitivity for some compounds
Co-elution difficult to detect

6. Explain the construction and working of thin layer chromatography with a neat
diagram
Thin layer chromatography (TLC) is a chromatography technique used to separate nonvolatile mixtures.[1] Thin layer chromatography is performed on a sheet of glass, plastic, or
aluminium foil, which is coated with a thin layer of adsorbent material, usually silica gel,
aluminium oxide, or cellulose. This layer of adsorbent is known as the stationary phase.
After the sample has been applied on the plate, a solvent or solvent mixture (known as the
mobile phase) is drawn up the plate via capillary action. Because different analytes ascend
the TLC plate at different rates, separation is achieved.[2]
Thin layer chromatography can be used to monitor the progress of a reaction, identify
compounds present in a given mixture, and determine the purity of a substance. Specific
examples of these applications include: analyzing ceramides and fatty acids, detection of
pesticides or insecticides in food and water, analyzing the dye composition of fibers in
forensics, assaying the radiochemical purity of radiopharmaceuticals, or identification of
medicinal plants and their constituents [3]
A number of enhancements can be made to the original method to automate the different
steps, to increase the resolution achieved with TLC and to allow more accurate quantitative
analysis. This method is referred to as HPTLC, or "high performance TLC".
Irreversibly adsorbed compounds not detected
Plate preparation
TLC plates are usually commercially available, with standard particle size ranges to improve
reproducibility. They are prepared by mixing the adsorbent, such as silica gel, with a small
amount of inert binder like calcium sulfate (gypsum) and water. This mixture is spread as a
thick slurry on an unreactive carrier sheet, usually glass, thick aluminum foil, or plastic. The
resultant plate is dried and activated by heating in an oven for thirty minutes at 110 C. The
around 0.5 2.0 mm for preparative TLC.[4]
Technique
Chromatogram of 10 essential oils coloured with vanillin reagent.
The process is similar to paper chromatography with the advantage of faster runs, better
separations, and the choice between different stationary phases. Because of its simplicity and
speed TLC is often used for monitoring chemical reactions and for the qualitative analysis of
reaction products.
To run a thin layer chromatography, the following procedure is carried out: [5]
A small spot of solution containing the sample is applied to a plate, about 1.5
centimeters from the bottom edge. The solvent is allowed to completely evaporate off,
otherwise a very poor or no separation will be achieved. If a non-volatile solvent was
used to apply the sample, the plate needs to be dried in a vacuum chamber.
A small amount of an appropriate solvent (eluent) is poured into a glass beaker or any
other suitable transparent container (separation chamber) to a depth of less than 1
centimeter. A strip of filter paper (aka "wick") is put into the chamber, so that its
bottom touches the solvent, and the paper lies on the chamber wall and reaches almost
to the top of the container. The container is closed with a cover glass or any other lid
and is left for a few minutes to let the solvent vapors ascend the filter paper and
saturate the air in the chamber. (Failure to saturate the chamber will result in poor
separation and non-reproducible results).
The TLC plate is then placed in the chamber so that the spot(s) of the sample do not
touch the surface of the eluent in the chamber, and the lid is closed. The solvent
moves up the plate by capillary action, meets the sample mixture and carries it up the
thickness of the absorbent layer is typically around 0.1 0.25 mm for analytical purposes and
plate (elutes the sample). The plate should be removed from the chamber before the
solvent front reaches the top of the stationary phase (continuation of the elution will
give a misleading result) and dried.
Separation Process and Principle
Different compounds in the sample mixture travel at different rates due to the differences in
solvent.[6] By changing the solvent, or perhaps using a mixture, the separation of components
(measured by the Rf value) can be adjusted. Also, the separation achieved with a TLC plate
can be used to estimate the separation of a flash chromatography column.[7]
Development of a TLC plate, a purple spot separates into a red and blue spot.
Separation of compounds is based on the competition of the solute and the mobile phase for
binding places on the stationary phase. For instance, if normal phase silica gel is used as the
stationary phase it can be considered polar. Given two compounds which differ in polarity,
the more polar compound has a stronger interaction with the silica and is therefore more
capable to dispel the mobile phase from the binding places. Consequently, the less polar
compound moves higher up the plate (resulting in a higher Rf value). [6] If the mobile phase is
changed to a more polar solvent or mixture of solvents, it is more capable of dispelling
solutes from the silica binding places and all compounds on the TLC plate will move higher
up the plate. It is commonly said that "strong" solvents (eluents) push the analyzed
compounds up the plate, while "weak" eluents barely move them. The order of
strength/weakness depends on the coating (stationary phase) of the TLC plate. For silica gel
coated TLC plates, the eluent strength increases in the following order: Perfluoroalkane
(weakest), Hexane, Pentane, Carbon tetrachloride, Benzene/Toluene, Dichloromethane,
Diethyl ether, Ethylacetate, Acetonitrile, Acetone, 2-Propanol/n-Butanol, Water, Methanol,
Triethylamine, Acetic acid, Formic acid (strongest). For C18 coated plates the order is
reverse. Practically this means that if you use a mixture of ethyl acetate and hexane as the
their attraction to the stationary phase, and because of differences in solubility in the
mobile phase, adding more ethyl acetate results in higher Rf values for all compounds on the
TLC plate. Changing the polarity of the mobile phase will normally not result in reversed
order of running of the compounds on the TLC plate. An eluotropic series can be used as a
guide in selecting a mobile phase. If a reversed order of running of the compounds is desired,
an apolar stationary phase should be used, such as C18-functionalized silica.
As the chemicals being separated may be colorless, several methods exist to visualize the
spots:
fluorescent analytes like quinine may be detected under blacklight (366 nm)
Often a small amount of a fluorescent compound, usually manganese-activated zinc
silicate, is added to the adsorbent that allows the visualization of spots under UV-C
light (254 nm). The adsorbent layer will thus fluoresce light green by itself, but spots
of analyte quench this fluorescence.
Iodine vapors are a general unspecific color reagent
Specific color reagents exist into which the TLC plate is dipped or which are sprayed
onto the plate.[8] [9] [10]
o
Potassium permanganate - oxidation
Bromine
In the case of lipids, the chromatogram may be transferred to a PVDF membrane and
then subjected to further analysis, for example mass spectrometry, a technique known
as Far-Eastern blotting.
Once visible, the Rf value, or retardation factor, of each spot can be determined by dividing
the distance the product traveled by the distance the solvent front traveled using the initial
spotting site as reference. These values depend on the solvent used and the type of TLC plate
and are not physical constants.
Applications
In organic chemistry, reactions are qualitatively monitored with TLC. Spots sampled with a
capillary tube are placed on the plate: a spot of starting material, a spot from the reaction
mixture, and a cross-spot with both. A small (3 by 7 cm) TLC plate takes a couple of minutes
Analysis
to run. The analysis is qualitative, and it will show if the starting material has disappeared,
i.e. the reaction is complete, if any product has appeared, and how many products are
generated (although this might be underestimated due to co-elution). Unfortunately, TLCs
from low-temperature reactions may give misleading results, because the sample is warmed
to room temperature in the capillary, which can alter the reactionthe warmed sample
analyzed by TLC is not the same as what is in the low-temperature flask. One such reaction is
the DIBALH reduction of ester to aldehyde.
UNIT III
INDUSTRIAL GAS ANALYZERS AND POLLUTION MONITORING INSTRUMENTS
1. Define paramagnetic property and state the types of paramagnetic oxygen

analyzer
Those gases which seek the strongest part of the field are called paramagnetic gases.
Eg:nitric oxide, chlorine dioxide.
2. What is meant by magnetic susceptibility?
Magnetic susceptibility is defined as the ratio of intensity of magnetization induced in
the substance by the field and its field strength.
3. List the methods of NO2analyzer.
1.colorimetry
2.chemi luminescence
3.use of CO laser
4. Laser Opto acoustic spectroscopy.
4. What is chemiluminesence?
The phenomenon of emission of radiation from a chemically excited species is known
as chemiluminescence. It results due to the formation of new chemical bonds .The atoms
in the excited states posses higher energy levels than the ground state & while returning
to the ground state they emit radiations.
5. Define thermal conductivity.
Thermal conductivity is defined as the quantity of heat transferred in unit time in a gas
between two surfaces 1 cm apart, when the temperature difference between the surfaces is
1oC
6. State the principle of thermal conductivity analyzers.
The preheated metal conducts electricity at a particular rate. If the gas is passed over the
heated metal, the gas absorbs the heat from the metal. This action reduces the temperature
of the metal, in turn the conductivity. This change in conductivity gives the measure of
the gas passed over the metal.
PART A
7. State the limitations of magnetic wind type instrument.
Hydrocarbons & other combustible gases react with the heated filament & causes
changes in temperature resulting in large errors. Therefore sample shouldnt contain these
substances in it.
2.photometric analysis
3.gold film semiconductor sensor
4. solid state sensor
5.lead acetate tape staining method
9. Give the types of filaments used for thermal conductivity analyzers
Pt,W,Co,Ni,Fe
10. State the limitations of magnetic force type instrument.
1.Entry of moisture will change instrument response. Suitable filter must be used at
the sample inlet.
2.Any change in temperature of the gases cause a corresponding change in its
magnetic susceptibility .So suitable circuit has to be employed to maintain sample
temperature as constant.
PART B
1. Explain with neat sketch the construction and working of Infra red gas analyzers
Introduction:
The Infra red 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 infra red 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.
8. List the types of H2S analyzer.

1.Electrochemical cell
Construction:
1. The IR gas analyzers consists of IR light source, a motor controlled chopper, a reference
cell, a sample cell and detector
2. The Motor controlled chopper is used to produce electric pulses rather than a continuous
beam of light
3. The reference cell consists of gases which do not absorb IR radiation
4. The sample cell has two openings. One for gas inlet and another is for gas outlet
5. The sample gas is made to pass through the sample continuously through these openings
6. An IR radiation detector is installed to capture the radiation passing out from the sample
cell and the reference cell
Working:
1. The IR radiation is passed through the chopper and pulses of radiation are produced
2. This radiation is passed through the sample cell and the reference cell
3. The reference cell do not absorb IR radiation and hence the intensity of IR radiation will
not change
4. The diatomic molecules present in the sample absorb the IR radiation and hence the
intensity of the radiation will be reduced
5. 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
Schematic of IR gas analyzer:
2.Explain in detail thermal conductivity sensors.

Introduction:
Thermal conductivity analyzer makes use of thermal conductivity principle to measure amount
of gas to be measured in the flu gas.
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
Principle:
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
6. This results in output Voltage and it is the measure of the sample gas concentration
3. Explain in detail how lead acetate tape staining is used to determine H2S in flu gas
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:
Introduction:
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.
2.
3.
4.
The H2S gas is humidified by acetic acid solution.

The humidified H2S is allowed pass through the lead acetate tape via a reaction window.
The H2S reacts with lead acetate tape and produces lead sulphide which is brown in color.
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.
4. What is Chemiluminescence? Explain how chemiluminescene can be used to determine

NO2 present in flu gas?
Chemiluminesence:
The phenomenon of emission of radiation from a chemically excited species is known as
chemiluminescence. It results due to the formation of new chemical bonds .The atoms in the
excited states posses higher energy levels than the ground state & while returning to the ground
state they emit radiations.
Construction:
Construction:
1.
2.
3.
4.
5.
NO2 is inert to O3 , Hence the NO2 converted into NO using appropriate catalyst and heat.
A O3 genereator is available to generate sufficient quantity of O3.
A reaction chamber is present where the NO and O3 reacts and produces light,
A Band pass filter is used to filter unwanted wavelengths
A photomultiplier tube is used to convert light into electric signals
Working:
1. The NO2 is converted into NO and it is allowed to react with O3 in a reaction chamber
2. When NO reacts with O3 it produces NO2 and light. This light is of unique wave length.
3. This light may contain multiple wavelengths and to remove unwanted wavelengths a
band pass filter is employed.
4. The filtered light is allowed to fall on photomultiplier tube, which converts the light into
appropriate electrical signals.
5. This electrical signal will give the measure of NO2 present in the flu gas.
Schematic of NO2 Analyzer:
5. Explain with a neat sketch the construction and working of Hartman Braun oxygen
Analyzer
Introduction:
Molecules can either be attracted or repelled by magnetic materials. Oxygen has two unpaired
electrons and it is paramagnetic in nature (i.e.) it will be attracted towards the magnetic field.
The magnetic field attracts the oxygen towards it. As the oxygen passes over a heated metal
towards the magnetic field, the flow of current varies in the metal.
Schematic of Hartman Braun oxygen Analyzer:
Construction:
1. The equipment consists of a round glass tube with a passage at its centre connecting two
sides.
2. A semi circular magnet is placed over the central passage of glass tube
3. A heated metal wire is wound over the central passage of glass tube
4. The metal wire is connected to one of the arms of the wheat stone bridge.
Principle:
1. When the air without oxygen is passed through the glass tube it will not be attracted
by the magnetic field and it will not pass through the central passage.
2. When the sample gas containing oxygen is passed through the tube, it gets attracted
towards the magnetic field and it passes through the central passage.
3. As the oxygen passes through the central passage, the heated platinum wire transmits
heat to the oxygen.
4. Since there is a change in the temperature of platinum wire, the current flow through
it changes, resulting in unbalanced bridge circuit.
5. The current reading showed in bridges galvanometer gives the measure of oxygen
present in the sample gas.
6. Explain with a neat sketch, magnetic force oxygen analyzers
Introduction:
Molecules can either be attracted or repelled by magnetic materials. Oxygen has two unpaired
electrons and it is paramagnetic in nature (i.e.) it will be attracted towards the magnetic field.
Principle:
When oxygen passes through a magnetic field along with a metal present in the magnetic field,
the magnetic susceptibility of the metal changes. This gives the measure of oxygen present in the
sample gas.
Working:
Construction:
Working:
1. When the sample gas containing oxygen is passed between the magnetic field, It gets
attracted by the magnetic field and it changes the magnetic susceptibility of the metal
sphere.
2. Due to the change in the magnetic susceptibility, the metal sphere twists.
3. As the sphere twists, the reflector attached to the sphere also moves.
4. The movement in the reflector gives a change in the light illumination from the lamp on
to the detector.
5. The light reflected from the reflector is gathered by a phototube and appropriate electrical
signals are generated.
6. The electrical signal is the measure of the oxygen present in the flu gas.
1. The equipment consists of two magnets, a metal sphere , a reflector , lamp and a photo
detector.
2. The magnetic field will be uneven and the metal sphere is suspended in a quartz wire.
3. The detector is connected to an electrical feedback circuit.
4. The sample is allowed to pass in between the poles of the magnet
UNIT IV
PH METERES AND DISSOLVED ANALYZERS
PART A
1. Define pH.
2. What is electrode slope?

Electrode slope is defined as the change in mill volt observed with every
tenfold change in concentration. For ammonia electrode the change in milli volts
should be in the range of -54 to -60 mV/decade, when the solution temperature is
between 20 to 25 oC
3. What are the limitations of glass electrode?
1. It gives accurate results when pH is up to 9.Above this it gives result
higher than the results given by electrode in same solution called alkali
error.
2. It cannot be used in strong alkaline solution.
3. Highly acid solution, concentrated alcoholic solution high temperature
and presence of ionized colloidal particles introduce errors when this electrode are
used.
4. Explain the principle of ion selective electrodes.
It is an electrochemical sensor that generates a millivolt potential when
immersed in a conducting solution. The magnitude of the potential is a function of the
logarithm of specific ion activity.
5. What are the limitations of hydrogen electrode?
1. It requires hydrogen-generating apparatus.
2. Care has to be taken to avoid poisoning of platinum black so that
efficiency is not ruined.
3. Hydrogen gas used is to be purified from all impurities and from
oxygen.
4. No rubber tubing is to be used for connections as rubber may contain
sulphur which contaminates the black & polythene or PVC tubing is to be used.
5. Hydrogen electrode cannot be used in presence of oxidizing agents
such as ferrous salts, very strong reducing agent and metallic salts which poison the
platinum black.
6. What is asymmetric potential?
pH of the solution refers to the hydrogen ion concentration or

hydroxyl ion in that solution.
When similar electrodes and similar solutions are placed on both sides of
glass electrode, potential obtained is never zero. The difference is generally
+2mV or even less called asymmetric potential. This is due to difference in the strain
on outer & inner surface of bulb.
8. What is bio-sensors?
Modification of ion-selective electrode, in which enzyme is immobilized
on the surface of the glass electrode behind a polymer membrane, which would result
in bio-molecule specificity. Such type of sensors is called bio-sensor.
9. State the advantages of ammonia electrode.
1. Ammonia electrode allows measurement not only of dissolved ammonia
in aqueous solution but also can be used to measure the ammonium ion after
conversion to ammonia or organic nitrogen after digestion of the sample.
2. The performance of the electrode is not affected by sample colour and
turbidity.
3. There is no interference from almost all anions, cations and dissolved
spices other than volatile amines.
10. State the application and disadvantage of bio-sensor.
Application:
1.It is used in medical fields.
2.used in food and beverage industries.
Disadvantages:
1.limited life time which restrict their commercial viability.
2.necessary improvements in their stability.
7. What is reference or calomel electrode?

The reference electrode against which the potential electrode of glass electrode is
measured is called calomel electrode.
PART B
1.
Explain with neat sketch the dissolved oxygen analyzer
Introduction:
Oxygen analyzers are widely used in industries to detect the amount of oxygen present in the
water in order to avoid corrosion in the metallic part of the boiler. The analyzer is based on
the katharometer or thermal conductivity.
The amount of oxygen in a closed space above water at a constant temperature depends upon
the oxygen content present in the water only and does not depend on gas above the water.
Schematic of dissolved oxygen analyzer:
Construction:
The dissolved oxygen analyzer consists of
Condensing section
Transmitting section
Analyzing section
The condensing section cools the sample water to be tested to 20 -25 0C. The sample water is
made to flow in a coiled tube around which cooled water is passed.
Principle:
The Transmitting section consists of flow regulator, which regulates the cool water to be
flowed into analyzing section.
The heart of the analyzer is analyzing section. The analyzing section terminals are connected
to the wheat stone bridge. The analyzing section consists of reference arm to which a
platinum wire is connected. This platinum wire is exposed to inert hydrogen gas. The other
platinum wire is made to expose to oxygen which is present above the water. This platinum
wire is connected to the measuring arm.
(i)
Balancing the Bridge:
Initially the bridge would not be balanced. To balance the bridge the amount of hydrogen
ions in the reference arm has to be adjusted. Until the balance is arrived, the sample should
not enter the measuring arm. The arm is subjected to a standard solution which produces a
constant oxygen rate. The sample is blocked from entering the contact tube by a water cock.
(ii)
Measuring dissolved oxygen:
Once the bridge is balanced, the water cock is opened and the sample water is allowed to
flow through the measuring arm. The contact tube picks up the oxygen gas and it changes the
temperature of the platinum wire placed in the contact tube and hence the current flows
through the bridge. The amount of current flow gives the measure of dissolved oxygen
present in the sample.
2. Explain with a neat sketch ammonia electrode
Introduction:
It is one of the most commonly used ion selective electrodes. The electrode uses a
hydrophobic gas-permeable membrane to separate the sample solution from the electrode
internal solution.
Principle:
Dissolved Ammonia on the sample solution diffuses through the membrane until the partial
pressure of ammonia is the same on both sides of the membrane. The partial pressure of
ammonia is proportional to its concentration.
Schematic of Ammonia Electrode:
Working:
Construction:
The ammonia electrode consists of outer body, inner body, sensing element and
ammonia selective membrane.
The inner body is filled with saturated Ammonium chloride. Since the solution is
saturated, the concentration of Ammonium will be fixed.
The ammonia sensing membrane allows only ammonium ions to diffuse through the
electrode.
An internal reference element is present, which acts as a reference electrode
producing constant potential.
The potential developed in ammonia electrode will be subtracted with the reference
electrode potential in order to achieve noise free and accurate value.
Working:
When ammonia diffuses through the membrane, it will dissolve in internal filling solution
and react reversibly with water as follows
3 + 2 +4 +
The relationship between ammonia, ammonium ion and hydroxide is given by
4 +
[3 ] =
Since the internal filling solution contains saturated solution of Ammonium chloride, we
arrive at a relation
= [3 ]Constant
The potential developed between the electrode sensing element and the reference electrode
varies with the changes in hydroxide level
= 0 log
[ ]
Since ammonia concentration is proportional to OH concentration we get

= 0 log
[3 ]
Disadvantages:
1. High electrode potential drift due to presence of water vapour movement in internal
filling solution
2. High noise.
3. Explain in detail the glass electrode
Introduction
Glass electrode is an ion selective electrode, sensitive, sensitive to H+ ions, so it is used to
measure H+ ion concentrations. It is used with other reference electrodes to generate a
potential difference.
Principle
A thin glass membrane is in contact with solutions of different H+ ion concentrations on its
two sides, a difference in potential is developed across the membrane. The magnitude of
potential difference depends on the difference in H+ ion concentration and hence the PH can
be calculated.
Schematic of Glass electrode
Where S is the slope factor
Construction
The glass electrode consists of the following parts
pH glass
outer electrolyte
Inner fill solution
Reference wire
Fill hole
Electrode (Silver chloride)
A brief construction of glass electrode is given below.

It is made up of a special glass of relatively low melting point and high electrical
conductivity, in the form of a sealed glass tube filled with 0.1 M Hcl
a platinum wire is inserted into it, to make electrical contact. The glass electrode is
represented as Pt; 0.1M HCL glass test H+ solution.
The H+ ion concentration inside the electrode is constant. When this electrode is
immersed into a solution of unknown H+, it becomes sensitive to the outside H+
concentration in the solution. Such sensitivity arises because of the difference
between the H+ ion concentration inside and outside.
A second electrode is necessary when measuring the electromotive force generated at
the electrode membrane of a glass electrode. This other electrode, paired with the
The reference electrode must have extremely stable potential. Therefore, it is
provided with a pinhole or a ceramic material at the liquid junction.
The Fill hole is used to fill the outer electrolyte.
Working:
The difference in pH between solutions inside and outside the thin glass membrane
creates electromotive force in proportion to this difference in pH.
This thin membrane is called the electrode membrane. Normally, when the
temperature of the solution is 30 , if the pH inside is different from that of outside
by 1, it will create approximately 60 mV of electromotive force.
The liquid inside the glass electrode usually has a pH of 7. Thus, if one measures the
electromotive force generated at the electrode membrane, the pH of the sample can
be found by calculation.
Advantages:
1. It can be used for the determination of PH of oxidizing agents, reducing agents
viscous madia, in presence of proteins which interfere the other electrode etc.
2. It can be used in the PH range of 0 to 12.
3. It can be used in colored, turbid and colloidal solutions.
4.
It is simple to operate.
Disadvantages:
1. Not used in ordinary potentiometer due to the high resistance of glass membrane.
glass electrode, is called the reference electrode
4. Explain in detail the calomel electrode
Introduction:
The Saturated calomel electrode (SCE) is a reference electrode based on the reaction between
elemental mercury and mercury(I) chloride.
A thin glass membrane is in contact with solutions of different concentrations on its two
sides, a difference in potential is developed across the membrane. The magnitude of potential
difference depends on the difference in concentration.
Schematic of Calomel Electrode:
Construction:
It consists of an outer glass tube fitted with a frit at the bottom to permit electrical
contact with the outside solution.
In side there is another tube, the bottom of which is packed with glass wool to allow
further electrical connection between the contents of the inner tube and the contents of
the outer tube.
Principle:
The inner tube is packed with a paste of mercury and mercurous chloride dispersed in
a saturated solution of potassium chloride
The electrode potential will depend on the concentration of the potassium chloride
and, thus, the electrode potential must be reported together with the potassium
chloride concentration. Thus, for the saturated calomel electrode the common
reference voltage is +0.244 V.
calomel electrode (SCE) and if the potassium chloride solution is 1 N, the electrode is
known as normal calomel electrode (NCE) while for 0.1 N potassium chloride
solution, the electrode is referred to as decinormal calomel electrode (DNCE).
Working:
The electrode reaction when the electrode acts as cathode is:
1/2 Hg2Cl2 + e- <---> Hg + Cl The reduction potentials of the calomel electrodes on hydrogen scale at 298K are as
follows:
Saturated KC1 0.2415 V
1.0NKC1
0.2800 V
0.1NKC1
0.3338 V
The electrode potential of any other electrode on hydrogen scale can be measured
when it is combined with calomel electrode.
The emf of such a cell is measured. From the value of electrode potential of calomel
electrode, the electrode potential of the other electrode can be evaluated.
Advantages:
1. The calomel electrode is more realistic and more robust and is one of the common
electrodes applied in corrosion analysis.
2. It is comparatively low-priced.
Disadvantages:
If potassium chloride solution is saturated, the electrode is known as saturated
1. The calomel electrode contains mercury, which poses health hazards
5. Explain in detail selective ion electrodes

Introduction:
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
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.
Principle:
The ion selective membrane selectively picks up certain ion to pass through it and creates a potential,
which can be measured using a voltmeter.
Schematic of Ion selective electrode:
Types of Ion Selective Membrane:

Glass membrane
Glass membranes
are
made
from
an
ion-exchange
type
of
glass
(silicate or chalcogenide). This type of ISE has good selectivity, but only for
several single-charged cations; mainly H+, Na+, and Ag+.
The glass membrane has excellent chemical durability and can work in very
aggressive media.
a voltmeter or pH meter.
A very common example of this type of electrode is the pH glass electrode.
Crystalline membrane
Crystalline membranes are made from mono- or poly crystallites of a single
substance. They have good selectivity, because only ions which can introduce
themselves into the crystal structure can interfere with the electrode response.
An example is the fluoride selective electrode based on LaF3 crystals.
Ion-exchange resins are based on special organic polymer membranes which
contain a specific ion-exchange substance (resin).
This is the most widespread type of ion-specific electrode.
An example is the potassium selective electrode, based on valinomycin as an
ion-exchange agent.
Construction:
These electrodes are prepared from glass capillary tubing approximately 2
millimeters in diameter, a large batch at a time.
Polyvinyl
chloride is
dissolved
in
a solvent and plasticizers (typically
phthalates) added, in the standard fashion used when making something out of
vinyl.
In order to provide the ionic specificity, a specific ion channel or carrier is
added to the solution; this allows the ion to pass through the vinyl, which
prevents the passage of other ions and water.
One end of a piece of capillary tubing about an inch or two long is dipped into
this solution and removed to let the vinyl solidify into a plug at that end of the
tube.
Using a syringe and needle, the tube is filled with salt solution from the other
end, and may be stored in a bath of the salt solution for an indeterminate
period.
For convenience in use, the open end of the tubing is fitted through a tight oring into somewhat larger diameter tubing containing the same salt solution,
with a silver or platinum electrode wire inserted.
New electrode tips can thus be changed very quickly by simply removing the
older electrode and replacing it with a new one.
Ion exchange resin Membrane
Working:
An ISE (is immersed in an aqueous solution containing the ions to be
measured, together with a separate, external reference electrode.
The electrochemical circuit is completed by connecting the electrodes to a
sensitive milli-volt meter using special low-noise cables and connectors.
A potential difference is developed across the ISE membrane when the target
concentration side
Advantages:
Inexpensive and simple to use
Wide concentration range
Unaffected by sample color or turbidity
Ideal for monitoring environmental pollution or water quality
Ideal for long term monitoring of changes in ion concentration
ions diffuse through from the high concentration side to the lower
6.
Explain Standard hydrogen electrode with a neat sketch
Introduction:
A Standard Hydrogen Electrode (SHE) is an electrode that scientists use for reference on all
half-cell potential reactions. The value of the standard electrode potential is zero, which
forms the basis one needs to calculate cell potentials using different electrodes or different
concentrations.
Construction and Working:

SHE is composed of a 1.0 M H+(aq) solution containing a square piece of platinized platinum
(connected to a platinum wire where electrons can be exchanged) inside a tube. During the
reaction, hydrogen gas is then passed through the tube and into the solution causing the
reaction:
2H+(aq) + 2e- <==> H2(g).
First an initial discharge allows electrons to fill into the highest occupied energy level of Pt.
As this is done, some of the H+ ions form H3O+ ions with the water molecules in the solution.
These hydrogen and hydronium ions then get close enough to the Pt electrode (on the
platinized surface of this electrode) to where a hydrogen is attracted to the electrons in the
metal and forms a hydrogen atom. Then these combine with other hydrogen atoms to create
H2(g). This hydrogen gas is released from the system. In order to keep the reaction going, the
electrode requires a constant flow of H2(g). The Pt wire is connected to a similar electrode in
which the opposite process is occurring, thus producing a charge that is referenced at 0 volts.
Other standard electrodes are usually preferred because the SHE can be a difficult electrode
to set up. The difficulty arises in the preparation of the platinized surface and in controlling
the concentration of the reactants. For this reason the SHE is referred to as a hypothetical
electrode.
Schematic of SHE:
UNIT V
ELECTRO MAGNETIC RESONANCE AND MICROSCOPIC TECHNIQUES
PART A
2. What is NMR?
It is the branch of spectroscopy in which the absorption of RF
nuclei of the sample subjected to magnetic field is studied.
radiation by
3. What are the types of mass spectrometer?

1.magnetic deflection mass spectrometers.
2. time of flight mass spectrometer.
3. radio frequency mass spectrometer
4. quadrapole mass analyzer
4. State the principle of mass analyzer.
The sample to be analyzed is first bombarded with an electron
beam to
produce ionic fragments of the original molecule .These ions are then sorted out by
accelerating them through electric field and magnetic field according to mass/charge ratio.
A record of numbers of different kinds of ions called mass spectrum.
5. State the limitations of quadrapole mass analyzer.
The performance of a mass filter depends on the quality of the quadrapole rods, the
stability of the applied voltage and the field characteristic at the two ends of the rods.
Especially, it is essential that the Quadra pole rod assembly is produced with super high
precision
6. State the advantages and disadvantages of time of flight mass spectrometer.
Advantages:
Speed, ability to record entire mass spectrum in one time. a conventional spectrometer
detects only one peak at a time. Its accuracy depends on electronic circuits than
mechanical alignment and on production of highly stable and uniform magnetic field.
Disadvantage:
Their poor resolution due to display on an oscilloscope.
1. What is the principle of radio frequency mass spectrometer?

The charged particles emerging from the ion source are all accelerated to the same
energy in an electrostatic field and then they pass through a system of RF electrodes. The
energy acquired by the ions in this process is a function of their specific mass/charge ratio.
7. What is meant by nuclear shielding?
The magnetic field at the nucleus is not equal to the applied magnetic field; electrons around
the nucleus shield it from the applied field. The difference between the applied magnetic
field and the field at the nucleus is termed the nuclear shielding.
9. What are the rules to determine Nuclear spin

If the number of neutrons and the number of protons are both even, then the nucleus
has NO spin.
If the number of neutrons plus the number of protons is odd, then the nucleus has a halfinteger spin (i.e. 1/2, 3/2, 5/2)
If the number of neutrons and the number of protons are both odd, then the nucleus has an
integer spin (i.e. 1, 2, 3)
10.
Draw the energy level of the nucleus with spin quantum number
PART B
1. Explain in detail the NMR Spectrometer
Introduction:
NMR spectrometer is the effective device used to analyze the structural components of
compound especially organic compounds.
Principle:
The nucleus of the atom will be spinning and it posses its own magnetic field and frequency.
At a particular magnetic field strength, the compound absorbs resonant frequency.
8. What is the principle of Electron spin resonance?

When the resonant frequency of the cavity matches with the frequency of the microwave
produced from the klystron, the incident microwave energy on the cavity will be absorbed
and the energy of the electron increases.
Construction:
A solution of the sample in a uniform 5 mm glass tube is oriented between the poles of a
powerful magnet, and is spun to average any magnetic field variations, as well as tube
imperfections.
Radio frequency radiation of appropriate energy is broadcast into the sample from an antenna
coil
A receiver coil surrounds the sample tube, and emission of absorbed rf energy is monitored
by dedicated electronic devices and a computer.
An nmr spectrum is acquired by varying or sweeping the magnetic field over a small range
while observing the rf signal from the sample.
An equally effective technique is to vary the frequency of the rf radiation while holding the
external field constant.
Working:
The NMR spectrometer can be operated in two ways
Keeping magnetic field constant and varying electric field
Keeping the electric field constant and varying the magnetic field
1. The Magnetic field is kept constant and the electric field frequency is increased.
Schematic of NMR spectrometer:
2. The lattice vibration of the nucleus will be at a frequency fn
3. If the frequency of the field is increased and if it is equal to fn resonance occurs.
4. At resonance, the low energy nucleus will absorb the EM radiation and excites to
higher states.
5. This can be graphically represented with ppm on x axis and energy of radiation on y
axis.
Introduction
SEM stands for scanning electron microscope. The SEM is a microscope that uses electrons
instead of light to form an image. Since their development in the early 1950's, scanning
electron microscopes have developed new areas of study in the medical and physical science
communities. The SEM has allowed researchers to examine a much bigger variety of
specimens.
Schematic of SEM:
Construction and Working

The SEM is an instrument that produces a largely magnified image by using electrons instead
of light to form an image. A beam of electrons is produced at the top of the microscope by an
electron gun. The electron beam follows a vertical path through the microscope, which is held
within a vacuum. The beam travels through electromagnetic fields and lenses, which focus
the beam down toward the sample. Once the beam hits the sample, electrons and X-rays are
ejected from the sample.
2. Explain with a neat sketch, the working of a scanning electron microscope
Detectors collect these X-rays, backscattered electrons, and secondary electrons and convert
them into a signal that is sent to a screen similar to a television screen. This produces the final
image.
Advantages
The scanning electron microscope has many advantages over traditional microscopes. The
SEM has a large depth of field, which allows more of a specimen to be in focus at one time.
The SEM also has much higher resolution, so closely spaced specimens can be magnified at
much higher levels. Because the SEM uses electromagnets rather than lenses, the researcher
has much more control in the degree of magnification. All of these advantages, as well as the
actual strikingly clear images, make the scanning electron microscope one of the most useful
instruments in research today.
3. Explain the construction and working of a Transmission electron microscope with a
neat sketch
Introduction:
A Transmission Electron Microscope (TEM) utilizes energetic electrons to provide
morphologic, compositional and crystallographic information on samples
Schematic of TEM:
Construction:
TEMs consist of the following components:
An electron source
Thermionic Gun
Electron beam
Electromagnetic lenses
Vacuum chamber
2 Condensers
Sample stage
Phosphor or fluorescent screen
Computer
A Transmission Electron Microscope functions under the same basic principles as an optical
microscope.
In a TEM, electrons replace photons, electromagnetic lenses replace glass lenses and images
are viewed on a screen rather than through an eyepiece.
Working:
A Transmission Electron Microscope produces a high-resolution, black and white image from
the interaction that takes place between prepared samples and energetic electrons in the
vacuum chamber.
Air needs to be pumped out of the vacuum chamber, creating a space where electrons are able
to move.
The beam passes through the solenoids, down the column, makes contact with the screen
where the electrons are converted to light and form an image.
The image can be manipulated by adjusting the voltage of the gun to accelerate or decrease
the speed of electrons as well as changing the electromagnetic wavelength via the solenoids.
The coils focus images onto a screen or photographic plate.
During transmission, the speed of electrons directly correlates to electron wavelength; the
faster electrons move, the shorter wavelength and the greater the quality and detail of the
image.
The lighter areas of the image represent the places where a greater number of electrons were
able to pass through the sample and the darker areas reflect the dense areas of the object.
These differences provide information on the structure, texture, shape and size of the sample.
To obtain a TEM analysis, samples need to have certain properties. They need to be sliced
thin enough for electrons to pass through, a property known as electron transparency.
Samples need to be able to withstand the vacuum chamber and often require special
preparation before viewing.
Types of preparation include dehydration, sputter coating of non-conductive materials,
cryofixation, sectioning and staining.
TEM Applications
A Transmission Electron Microscope is ideal for a number of different fields such as life
sciences, nanotechnology, medical, biological and material research, forensic analysis,
gemology and metallurgy as well as industry and education.
TEMs provide topographical, morphological, compositional and crystalline information.
The electrons then pass through multiple electromagnetic lenses. These solenoids are tubes
with coil wrapped around them.
The images allow researchers to view samples on a molecular level, making it possible to
analyze structure and texture.
This information is useful in the study of crystals and metals, but also has industrial
applications.
Technology companies use TEMs to identify flaws, fractures and damages to micro-sized
objects; this data can help fix problems and/or help to make a more durable, efficient product.
Colleges and universities can utilize TEMs for research and studies.
Although electron microscopes require specialized training, students can assist professors and
learn TEM techniques.
Students will have the opportunity to observe a nano-sized world in incredible depth and
detail.
Advantages
A Transmission Electron Microscope is an impressive instrument with a number of
advantages such as:
TEMs offer the most powerful magnification, potentially over one million times or more
TEMs have a wide-range of applications and can be utilized in a variety of different

scientific, educational and industrial fields
TEMs provide information on element and compound structure
Images are high-quality and detailed
TEMs are able to yield information of surface features, shape, size and structure
They are easy to operate with proper training
Disadvantages
Some cons of electron microscopes include:
TEMs are large and very expensive
Laborious sample preparation
Potential artifacts from sample preparation
Operation and analysis requires special training
TEMs can be used in semiconductor analysis and production and the manufacturing of
computer and silicon chips.
Samples are limited to those that are electron transparent, able to tolerate the vacuum
chamber and small enough to fit in the chamber
TEMs require special housing and maintenance
Images are black and white
A Transmission Electron Microscope requires constant upkeep including maintaining

voltage, currents to the electromagnetic coils and cooling water.
4. Explain with neat sketch, electron spin resonance spectrometer
Introduction:
Electron Spin resonance spectroscopy is based on the absorption of microwave radiation by
an unpaired electron when it is exposed to a strong magnetic field. Species that contain
unpaired electrons (namely free radicals, odd-electron molecules, transition metal complexes,
rare earth ions, etc.) can therefore be detected by ESR.
Principle:
When the resonant frequency of the cavity matches with the frequency of the microwave
produced from the klystron, the incident microwave energy on the cavity will be absorbed
and the energy of the electron increases.
Schematic of Electron spin resonanace:
Construction:
Electron microscopes are sensitive to vibration and electromagnetic fields and must be
housed in an area that isolates them from possible exposure.
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.
Attenuator:
The power level of microwaves will be very high. To reduce the power level of the
microwave, the attenuator is used.
Cavity:
The sample is mounted in the microwave cavity
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 sample is mounted in the electric field nodal plane, but at a maximum in the magnetic
field.
Working:
Klystron Tube:
Since the cavity length is not adjustable but it must be exactly one wavelength, the
spectrometer must be tuned such that the klystron frequency is equal to the cavity resonant
frequency. The tune-up procedure usually includes observing the klystron power mode. That
is, the klystron reflector voltage is swept, and the diode current is plotted on an oscilloscope
or other device. When the klystron frequency is close to the cavity resonant frequency, much
less power is reflected from the cavity to the diode, resulting in a dip in the power mode. The
"cavity dip" is
centered on the power mode using the coarse mechanical frequency adjustment with
the reflector voltage used to fine tune the frequency.
5. Explain the conceptual aspects of Mass spectrometer with necessary diagram
Atoms can be deflected by magnetic fields - provided the atom is first turned into an
ion. Electrically charged particles are affected by a magnetic field although electrically
neutral ones aren't.
The sequence is :
Stage 1: Ionisation
The atom is ionised by knocking one or more electrons off to give a positive ion. This is
true even for things which you would normally expect to form negative ions (chlorine,
for example) or never form ions at all (argon, for example). Mass spectrometers always
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.
work with positive ions.
Stage 2: Acceleration
The ions are accelerated so that they all have the same kinetic energy.
The ions are then deflected by a magnetic field according to their masses. The lighter
they are, the more they are deflected.
The amount of deflection also depends on the number of positive charges on the ion - in
other words, on how many electrons were knocked off in the first stage. The more the
ion is charged, the more it gets deflected.
Stage 4: Detection
The beam of ions passing through the machine is detected electrically.
A full diagram of a mass spectrometer
Ionisation
Stage 3: Deflection
The vaporised sample passes into the ionisation chamber. The electrically heated metal
coil gives off electrons which are attracted to the electron trap which is a positively
charged plate.
The particles in the sample (atoms or molecules) are therefore bombarded with a stream
of electrons, and some of the collisions are energetic enough to knock one or more
electrons out of the sample particles to make positive ions.
Most of the positive ions formed will carry a charge of +1 because it is much more
difficult to remove further electrons from an already positive ion.
These positive ions are persuaded out into the rest of the machine by the ion repeller
which is another metal plate carrying a slight positive charge.
Acceleration
The positive ions are repelled away from the very positive ionisation chamber and pass
through three slits, the final one of which is at 0 volts. The middle slit carries some
intermediate voltage. All the ions are accelerated into a finely focused beam.
Deflection
Different ions are deflected by the magnetic field by different amounts. The amount of
deflection depends on:
the mass of the ion. Lighter ions are deflected more than heavier ones.
the charge on the ion. Ions with 2 (or more) positive charges are deflected more than
ones with only 1 positive charge.
These two factors are combined into the mass/charge ratio. Mass/charge ratio is given
the symbol m/z (or sometimes m/e).
For example, if an ion had a mass of 28 and a charge of 1+, its mass/charge ratio would
be 28. An ion with a mass of 56 and a charge of 2+ would also have a mass/charge ratio
of 28.
In the last diagram, ion stream A is most deflected - it will contain ions with the
smallest mass/charge ratio. Ion stream C is the least deflected - it contains ions with the
greatest mass/charge ratio.
It makes it simpler to talk about this if we assume that the charge on all the ions is 1+.
Most of the ions passing through the mass spectrometer will have a charge of 1+, so
that the mass/charge ratio will be the same as the mass of the ion.
Assuming 1+ ions, stream A has the lightest ions, stream B the next lightest and stream
C the heaviest. Lighter ions are going to be more deflected than heavy ones.
Detection
Only ion stream B makes it right through the machine to the ion detector. The other
ions collide with the walls where they will pick up electrons and be neutralised.
Eventually, they get removed from the mass spectrometer by the vacuum pump.
When an ion hits the metal box, its charge is neutralised by an electron jumping from
the metal on to the ion (right hand diagram). That leaves a space amongst the electrons
in the metal, and the electrons in the wire shuffle along to fill it.
A flow of electrons in the wire is detected as an electric current which can be amplified
and recorded. The more ions arriving, the greater the current.
6. Explain quadrapole Mass spectrometer
Introduction:
A Quadrupole is a mass analyzer that uses an electric field to separate ions.
Principle:
The ions, when passed in between the electric field, it will get deflected. The amount of
deflection is based on its mass.
Construction and Working:
The Quadrupole consists of 4 parallel rods/ poles, where adjacent rods have opposite
voltage polarity applied to them.
The voltage applied to each rod is the summation of a constant DC voltage (U) and a
varying radio frequency (Vrfcos(wt)),
where w = angular frequency of the radio frequency field.
The electric force on the ions causes the ions to oscillate/orbit in the area between the 4
rods, where the radius of the orbit is held constant.
The ion moves in a very complex motion that is directly proportional to the mass of the
ion, voltage on the quadrupole, and the radio frequency.
The ions will remain orbiting in the area between the poles with no translation along the
length of the poles unless the ions have a constant velocity that is created as the ions
enter the quadrupole.
Before entering the analyzer, the ions travel through a potential of a certain voltage,
usually created by ring electrode, in order to give the ions a constant velocity so they
can transverse along the center of the quadrupole.
While in the quadrupole, the trajectories of the ions change slightly based on their
masses.
Ions of specific mass have a certain frequency by which they oscillate. The greater the
mass, the greater the frequency.
A certain limit is associated with each quadrupole and it selects ions which are within
the desirable frequency range.

Unit I Colorimeter and Spectrophotometers: 2. What Is Meant by Flame Emission Spectrometry?

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Unit I Colorimeter and Spectrophotometers: 2. What Is Meant by Flame Emission Spectrometry?

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2. What is meant by flame emission spectrometry?

3. State Beers law.

5. What is analytical instrumentation?

1. List any four elements used in spectrophotometers.

6. What is electromagnetic radiation?

vacuum. It may be considered as discrete packets of energy called photons.

Draw the spectrum of EM regions.

9. State Lamberts law.

Explain with necessary diagrams the instrumentation involved in uv visible

schematic diagram of a double-beam UV-Vis. spectrophotometer

The linear photodiode array is an example of a multichannel photon detector. These

The diffraction grating and the slit

wavelengths through into the rest of the spectrometer.

The rotating discs

It is following the red path in the diagram:

in the wavelength range we are interested in (200 - 800 nm).

The detector and computer

An absorbance of 0 at some wavelength means that no light of that particular

Explain the construction and working of FTIR?

Figure 2. Block diagram of an FTIR spectrometer

Figure 3. Schematic of the Michelson interferometer

A typical Michelson interferometer consists of two perpendicular mirrors and a beamsplitter.

Fourier Transform of Interferogram to Spectrum

Figure 4. (a) Interferogram of a monochromatic light; (b) its spectrum

where A is the measured absorbance, a

is a wavelength-dependent absorptivity coefficient,

is the wavelength-dependent molar absorptivity coefficient with units of M -1 cm-1.

* N * A * dz. Then, the

Integrating this equation from z = 0 to z = b gives:

maximum. Taking an infinitesimal slab, dz, of sample:

Since N (molecules/cm3) * (1 mole / 6.023x1023 molecules) * 1000 cm3 / liter = c

Limitations of the Beer-Lambert law

(moles/liter) and 2.303 * log(x) = ln(x), then

Chennai University .net

Figure 1. The Basic System of a Flame Atomic Absorption Spectrometer

Figure 2. The Hollow Cathode Element Lamp.

The analytical apparatus

converted into a dry aerosol which is volatilized in the flame.

Construction and working:

into a spray chamber.

separation (separated flame), or aerodynamically, by surrounding the flame with a

Liqid _Solid (LSC)

3. What is Retention Time?

Flame Photometric Detectors

Sample injection port

water and other impurities.

entering the column.

specific and non-destructive.

emissions monitoring in stationary or portable instruments.

Description of a generic detector

Techniques of Ion Exchange Chromatography:

2. Preparation of Ion Exchange

3. Washing of Ion Exchangers

Figure 2: Ion exchange chromatography

6. Development an Elution of bound ions

5.Explain the construction and working of High Pressure Liquid Chromatography

Schematic of an HPLC instrument

Co-elution difficult to detect

Irreversibly adsorbed compounds not detected

Potassium permanganate - oxidation

the DIBALH reduction of ester to aldehyde.

1. Define paramagnetic property and state the types of paramagnetic oxygen