Analytical Instruments

Reference Text: R S Khandpur

“Handbook of Analytical Instrumentation”

1
 Sources of infrared radiation
(Black body sources)
Globar Rod:
•Silicon Carbide rod with +ve temp coefficient
of resistance.
•Length= 5cm, Diameter=0.5cm
•Heated to 1500K
•Water cooling is required to prevent burnout.
•As Temp increases resistance increases. Hence
voltage should also be increased. So variable
transformer is used.
 Sources of infrared radiation
Nernst Lamp: (ZrO2, Y2O3)
Small rod of oxides of Zirconium and
Yttrium.
•-ve temp coefficient.
•Cylindrical in shape.
•Length= 20-30mm, d=1 to 2mm.
•Pt leads are sealed at the ends for passage of
current.
•Temp 1500 to 20000C
•Must be heated externally to dull-red hot
because it is non-conducting when cold.
 Sources of infrared radiation
•Nichrome Strip:
•Less energy than Globar or Nernst filament.
•Simple and reliable in operation
•Tightly wound Nichrome wire heated by
passing current.
•Temp: 800 to 9000C
•Requires no water cooling.
Optical Components used in Spectroscopy
 Entrance Slit: Purpose is to provide rectangular
optical image.
 Collimating Mirror or lens: Purpose is to produce
parallel beams of radiation, it overcomes diffraction.
 Prism or Grating: Disperses radiation into its
component wavelengths.
 Focusing Mirror or lens: Reforms image from slit
onto focal plane.
 Exit Slit: Isolates Spectral Band. 5
 Optical Components used in Spectroscopy

Diffraction Grating Monochromator Prism type Monochromator

6
 IR Detectors
Thermal, radiant energy  Electrical
energy
Detectors
Thermal Detectors Quantum Detectors

Eg: Thermocouple Eg: Intrinsic type

Thermopile - Solid state Photo Detectors

Bolometer Extrinsic type

Pneumatic - Photoconductive cell

detector
(Golay)
Pyroelectric 7
 Features
Thermal Detectors Quantum
Detectors

 Responsitivity with little  Responsitivity is

wavelength
dependence on wavelength.
dependent.
 Operation at room temperature.  Cooling is normally used.

 Slow response and  Fast response high

low detectivity. and detectivity.

8
 Quantum Detectors
Solid state Photo Detectors
 Principle:- Photo electric effect
 Materials:-Cadmium-Mercury-
Telluride(CMT) or
Indium Antimonide (InSb)
 Sensitivity:- 2-6 μ (InSb)
10-12 μ
(CMT)
Photoconductive cell
 Principle:- Electrical resistors,
which decrease in resistance in
relation to the intensity of light
striking there surface.
 Materials:-Semiconductors
like Lead sulphide or Lead
telluride
 Sensitivity:- 3.5 μ (Lead sulphide)
6 μ (Lead telluride)
9
 Thermal Detectors
1) Thermocouple
 Principle:- In these detectors, the signal originates from a
potential difference caused by heating a junction of the
unlike metals by the infrared beam, while the other
junction is kept at constant temperature.

10
 Thermal Detectors
2) Thermopiles
 Principle:- It is possible to increase the output voltage by
connecting several thermocouples in series. This
arrangement is referred to as Thermopiles.

11
 Thermal Detectors
3) Bolometer
Principle:- It provides an electrical signal as a result of the
variation in resistance of a conductor with temperature.

12
 Thermal Detectors
4) Pneumatic detector (Golay Detector)
Principle:- It measures the intensity of IR radiation by the
expansion of a gas filled in its chamber, upon heating.

Light source

Photocell

13
 Thermal Detectors
5) Pyroelectric detector
 Principle:- Pyroelectric effect
 In a pyroelectric detector which is composed of a pyroelectric
material, a change in temperature due to the application of IR
radiation creates a change in polarization . Such a crystal will
create an accumulation of charge and this charge is collected by
electrodes on the crystal.
 ie, by connecting 2 electrodes to the crystal, the pyroelectric
detector can act as a capacitor and the resulting voltage =
charge / crystal capacitance, V= Q/C.
 The detector will also ignore the effects of background
radiation. Pyroelectric detectors are commonly used in FTIR
spectrometers.
14
 Thermal Detectors
5) Pyroelectric detector
Commonly used crystal material for pyroelectric detector
is

Tri Glycine Sulphate (TGS).

15
 Thermal Detectors
5) Pyroelectric detector
 The very small electrical charges are generally converted within the
detector housing to convenient signal voltages by use of extremely
low noise and low leakage Field Effect Transistors (FET).

16
Response Times of Detectors
Detector Response Time

Photoconductive Cells 0.5ms

Golay Cell 4ms

Bolometer 4ms

Thermocouple 15-60ms
 FTIR Spectrometer
(Fourier Transform Spectrometer)
 Fourier Transforms
 Fourier transform defines a relationship between a
signal in time domain and its representation in
frequency domain.
 Being a transform, no information is created or lost in
the process, so the original signal can be recovered
from the Fourier transform and vice versa.
 The Fourier transform of a signal is a continuous
complex valued signal capable of representing real
valued or complex valued continuous time signals.
 What is a FTIR Spectrometer?
 FTIR (Fourier Transform InfraRed) spectrometer obtains an
infrared spectra by first collecting an interferogram of a
sample signal using an interferometer, then a
performs Fourier Transform on the
spectrum.
interferogram to obtain the

 An interferometer is an instrument that uses the technique of

superimposing (interfering) two or more waves, to detect
differences between them. The FTIR spectrometer uses a
Michelson interferometer.
 FT-IR Spectrometer
Principle:
Michelson’s Interferometer

21
 Schematic diagram of a Basic Optical Components of
Michelson interferometer for FTIR the Interferometer
(Principle)
 Interferometer
 Monochromatic radiation entering the interferometer is
split into 2 beams having two different path lengths by
Beam splitter.
 When beams A and B recombine, an interference pattern
is produced.
 A detector measures the intensity variations of the exit
beam as a function of path difference.
 When two beams are in phase at beam splitter, maximum
intensity will reach detector.
 Interferometer
 When two beams are out of phase intensity will
be minimum.
 When mirror M2 moved uniformly, Detector output will be
a sine wave.
 Amplitude of signal will depend the intensity of
on incoming radiation.
 Frequency is determined by
 Translation velocity of M2

 Wavelength of incoming
radiation
FTIR Spectrometer–Block Diagram
 Components of FTIR
1) IR Source (Glowbar)

2) Interferometer

3) Sample cell

4) Detector (Pyroelectric Detector)

5) Computer

6) Recorder or Plotter
Double Beam (Optical Null)
IR Spectrophotometers
Double Beam (Ratio Recording) IR
Spectrophotometer
Double Beam (Ratio Recording) IR
Spectrophotometer (Optical Diagram)
Double Beam (Ratio Recording) IR
Spectrophotometer
•Sector mirror when it rotates by 10rpm then the
modulation frequency is 20Hz.
• Detector-Thermocouple generates 0.5uV voltage.
 Advantages of FTIR
• Very high resolution (< 0.1 cm –1 )
Resolution governed by distance movable mirror travels

• Very high sensitivity (nanogram quantity)

can be coupled with GC analysis (–> measure IR spectra in gas-phase)

• High S/N ratios - high throughput

Few optics, no slits mean high intensity of light

• Rapid (<10 s)

• Reproducible and

• Inexpensive
 Applications
 Drug Analysis
 Fiber Analysis
 Paint Chip
Analysis
 Ink Analysis
 Paper Analysis
 Biological Analysis
 Flame Photometry
•According to the quantum theory, the energy states
of an atom or molecule are sharply defined and any
change from one state to another therefore, requires a
sharply defined quantity of energy.
•When radiation falls on a material, or the material is
supplied with extra energy in some form, some part
of the energy is taken up by the material and results
in altering the state of the atoms or molecules of
which it is composed. The atoms or molecules of the
material are promoted to higher energy states.
 Flame Photometry
•However, the higher energy states are rather
unstable. The particles at the higher energy levels
tend to lose the extra energy and return to the
original level or ground state, either by undergoing
a chemical reaction or by dissipating the energy as
heat, or by emitting the energy as radiation.
•If it loses all or part of the energy as radiation, it
will emit photons of an energy corresponding to
difference between two energy levels. Since the
levels are clearly defined for a given atom, the
radiation will be emitted at clearly defined
frequencies only.
 Flame Photometry
•The frequencies are shown up as bright lines if
the emitted light is dispersed as a spectrum.
•By measuring the wavelength of the emission,
one can find out what atoms are present.
•Also, by measuring the intensity of the
emission, one could compute the concentration
of the element.

E  h  hc / 
Flame Photometry
 Flame Photometry
Flame photometry offers the following
advantages:
• The technique is very rapid. It does not require
any chemical preparation except preparing a
solution of suitable concentration.
• The method is most useful for analysis of some
elements, which are difficult to measure by
other methods.
• The technique is most suited to analytical
problems, in which a large number of samples
of similar types are to be measured.
• The method is quite cheap as it does not
require any other expensive reagents.
 Flame Photometry
In addition, the analysis of alkali and alkaline
earth metals by flame photometry has the
following two major advantages:
• Their atoms reach the excited state at a
temperature lower than that at which most
other elements are excited.
• Their characteristic wavelengths are easily
isolated from those of most other elements
due to wide spectral separation.
 Basic Flame Photometer
•A solution of the sample to be analysed
is prepared. A special sprayer operated by
compressed air or oxygen is used to introduce
this solution in the form of a fine spray
(aerosol) into the flame of a burner operating
on some fuel gas, like acetylene or hydrogen.
•Conversion of sample solution into an aerosol
by atomiser does not bring about any chemical
change in the sample. However, the heat of the
flame which vaporises sample constituents,
molecules and ions of the sample species are
decomposed and reduced to give atoms.
 Basic Flame photometer
The heat of the flame causes excitation of some
atoms into higher electronic states. Excited
atoms revert to the ground state by emission of
high energy of characteristic wavelength.
The radiation of the element produced in the
flame is separated from the emission of other
elements by means of light filters or a
monochromator.
 Basic Flame Photometer
•The intensity of the isolated radiation is
measured from the current it produces when it
falls on a photocell. The measurement of
current is done with the help of a
galvanometer, whose readings are proportional
to the concentration of the element.
•After carefully calibrating the galvanometer
with solutions of known composition and
concentration, it is possible to correlate the
intensity of a given spectral line of the
unknown sample, with the amount of the same
element present in a standard solution.
Emission intensity Vs Concentration
Construction details of flame
photometer
Emission System:
It consists of the following:
(i) Fuel gases and their regulation:
comprising the fuel reservoir, compressors,
pressure regulators and pressure gauges.
(ii) Atomizer: consisting, in turn, of the
sprayer and the atomisation chamber, where
the aerosol is produced and fed into the
flame.
(iii) Burner: receives the mixture of the
combustion gases.
(iv) Flame: the true source of emission.
 Basic Flame Photometer
•Optical System: It consists of the optical
system for wavelength selection (filters or
monochromators), lenses, diaphragms, slits,
etc.
•Recording System: It includes detectors like
photocells, phototubes, photomultipliers, etc.,
and the electronic means of amplification,
measuring and recording.
Block Diagram
 Emission System

a) Fuel gases and their regulation:

•Pressure Regulators
•Flow meters
•Fuel supply
•Oxygen supply
•Air supply
 Fuel gases and their regulation
Pressure Regulators:
•For steady emission reading, flame should be
steady and free from flickering.
•To achieve this, the air or oxygen and fuel
pressure is maintained constant during the
operation of the instrument.
•Suitable pressure gauges are therefore,
provided in the instrument to indicate the
pressure actually present in the line.
•A 25-lb gauge for the oxygen or air supply and
a 10-lb gauge for the fuel are generally used.
 Fuel gases and their regulation
Pressure Regulators:
•Pressure regulators are usually followed by
needle valves for control of flow.
•Gauges provided with the regulators are often
not sensitive enough to detect small changes of
pressure, which have profound effect on the
flame photometer operation.
•Therefore, narrow range pressure regulators
and manometers are installed in the line, in
order to observe small changes in pressure or
gas flow.
 Fuel gases and their regulation
Flow meter:
•A flow meter may be inserted in the line from the
gas reservoir to the atomizer in order to detect
any clogging of the orifice.
•For the same flow rate, an appreciable change in
the gauge pressure indicates partially clogged
orifice.
•By controlling the individual flow rates of the
fuel and oxygen, the operator can choose various
fuel-oxygen mixtures ranging from lean flame
mixtures to fuel-rich types of flames.
•The flow rates usually vary from 2 to 10 Cu. ft/h.
 Fuel gases and their regulation
Fuel Supply:
•The fuel gas normally used in flame
photometry is the acetylene gas, which is
commercially available in cylinders of various
sizes.
•Cylinder acetylene consists of acetylene gas
dissolved in acetone, which in turn is absorbed
on a porous filling material.
•Consequently, after the flame is lit, it should be
allowed to burn several minutes before
adjustments are attempted in order to vent the
excess acetone initially present in the vapour
phase.
 Fuel gases and their regulation
Fuel Supply:
•Consumption of acetylene ranges from 1 to 5 s
Cu. ft/h. The other fuels used in flame
photometry are propane, butane and hydrogen.
•When the available gas pressure is less or is
variable, a booster pump is necessary.
• The pump generally used is a motor-driven
diaphragm pump, which delivers the gas to the
burner at the required volume and pressure.
 Fuel gases and their regulation
Oxygen Supply:
•Oxygen from cylinders should be supplied to the
burner through a regulator capable of delivering
approximately 12 Cu. ft/h. at a pressure of 12–15
lb per sq. inch.
Air Supply:
•Supplied from a cylinder of compressed air or
from an air compressor through a tank held at
about 10 lb/sq. inch.
•When compressors are used, the air should be
filtered through glass wool. Consumption of air is
approximately 10 Cu. ft/h for an acetylene-air
flame.
 Emission System: (b) Atomizer
•The usual method is to prepare solutions of known
concentrations and to spray these into the flame,
using some form of aerosol production.
•Use of an aerosol permits the distribution of the
same throughout the body of the flame, rather than
its introduction at a single point. In flame
photometry, the name atomizer is given to a system
which is used to form aerosol by breaking a mass of
liquid into small drops.
•This little device is responsible for introducing the
liquid sample into the flame at a stable and
reproducible rate. The atomizer must not be
attacked by corrosive solutions.
 Atomizer
Two types of atomisers which are in common
use are:
(i) Those which introduce the spray into a
condensing chamber and into the flame by
the air of the combustible gas air mixture.
Large droplets are removed in the
condensing chamber.
(ii) Those in which the sample is introduced
directly into the flame (i.e. the atomiser
and the burner are an integral unit).
 Atomizer
(i) Discharge type atomiser:
Atomizer: Discharge type atomiser:
•Consists of two capillary tubes sealed into the
walls of a glass chamber in such a manner that
their bores are perpendicular to each other.
•The sample solution is poured into a funnel or
drawn up from a container and is atomized by
the blast of air from the tip of the other
capillary.
•However, the atomized stream is composed of
coarse spray with large droplets, which
condense on the walls of the chamber and
helical tube leading to the burner.
Atomizer: Discharge type atomiser:
•The condensate flows down to the waste drain.
The smaller droplets, in the form of a virtual fog
are carried by the air stream into the burner,
where it is mixed with the burner gases and
carried into the region of active combination.
•Two removable hypodermic needles of stainless
steel or glass are commonly used. With this type
of atomiser, the consumption tilt sample is
comparatively high and ranges between 4 and
25 ml of solution per minute. Of this amount
only 5% actually reaches the flame.
Atomizer: Discharge type atomiser
•The sensitivity of this type of atomiser can be
markedly increased by using a chamber which is
heated by an electric heater placed around its
walls.
•This hastens the process of vaporisation of the
solvent and produces an aerosol of very fine
particles, all of which are swept into the burner.
•In this case, sample consumption is only 0.2–
0.6 ml/min and a substantial portion is carried
directly into the burner to yield a much higher
sensitivity.
Atomizer: Integral Burner type
 Atomizer
(ii) Integral Burner type Atomiser:
•In the sample solution is introduced directly
into the flame. The unit made of glass or metal
is constructed of two concentric tubes.
•The sample solution is drawn through the
innermost tube by the passage of oxygen
through the orifice of the middle annulus. At the
tip of the inner sample capillary, the liquid is
sheared off and dispersed into droplets.
Atomizer: Integral Burner type atomiser
•The outer annulus supplies the combustible gas
to the flame. The body of the unit is machined
from brass and the capillary for solution intake
is of palladium.
•All droplets, both large and small, are
introduced directly into the flame. Sample
consumption is between 0.8 and 2 ml/min.
 Atomizer
•Periodic cleaning of the atomiser and capillary
tube is necessary as blocked atomiser will give
unstable reading, intermittent reading and low
sensitivity.
•Cleaning is usually done by flushing with
copious amount of distilled water. If blockage
occurs, the atomiser is removed from its
reading and flushed with dry air or cleaned it
by using thin wire.
 Emission System: (c) Burner
The burner brings the fuel, oxidant and sample
aerosol together, so that they may react safely
and produce a good flame. Burners used in
flame photometry must fulfil the following
conditions:
(i) Supply fuel and oxygen at constant pressure
to enable the shape, size and temperature of the
flame will remain constant.
(ii) Assist in a perfect distribution of the mixture
of gases and the aerosol, which carries the
atomised solution under analysis.
 Burner
(iii) Have a tip of suitable shape to produce a
symmetrical flame and ensure a homogeneous
flow and distribution of the gases, avoiding a
strike back in the burner from accidental
fluctuations in the feed system.
(iv) Prevent condensation of the aerosol in the
stem of the burner, which would reduce the
effective quantity of the sample brought into
the flame.
 Emission System: (d) Flame
The flame is the most important part of the
flame photometer, since it forms the source in
which the light radiations, characteristic of the
elements under analysis, are produced. The
flame performs the following functions:
(i) It converts the constituents of the sample to
be analysed from the liquid or solid state into
gaseous state.
(ii) Decomposes these constituents into atoms
or simpler molecules.
(iii) Excites the resulting atomic or molecular
species to emit light radiations.
 Flame
•In order to produce accurate results, the
flame must be very stable. If its temperature
or structure shows a change over a period of
time, the emission produced by a given
sample will also change.
•The flame temperature must be high
enough to excite the atoms to higher energy
levels, so that emission may take place.
Flame
 Optical System
•Filters
•Monochromators
and other components like Reflectors,
Condensing lenses etc.
 Detectors
Photosensitive Detectors:
The photo detectors are either of
photovoltaic or photo-emissive type or any
one of the photo detector types.
 Recording System
Amplifiers for Phototube circuit:
•The current from the phototube is generally so
small that it is necessary to amplify it, quite often
by a factor of 105 to 106. The amplification of a
direct current can be achieved with the help of
electronic circuits with high input impedance.
•In order to reduce the rapid and random
fluctuations, a long enough time constant is
incorporated in the amplifier circuit to aid in the
integrating process, and thus the metre is
rendered unresponsive to sudden flashes of light
in the flame. Usually, the time constant is chosen
to be approximately 1 s.
Amplifiers for Phototube Circuit:
•To overcome the problem of drift of DC
amplifiers, the light from the flame can be
interrupted, and the photoelectric current
can be amplified by an AC amplifier.
•A synchronously rotating disc is placed
before the flame and the photoelectric
current is thus convened to a chopped DC
signal, which can then be amplified by an AC
amplifier.
Amplifiers for PMT:
Since PMT gives larger current amplifier
gain of the amplifier needed is around 10
only.
 Types of Flame Photometers
(i)Single beam
(ii) Double beam
(iii) Recording type
 Single Beam Flame Photometer

•B&A are coarse and fine sensitivity controls.

•The photoelectric current generated in the
photocell by light is given to the galvanometer.
 Double Beam Photometer

•It has second light path emitted by an internal standard

element. Each beam if focused on a photoelectric cell.
•The circuit is so arranged that the photoelectric currents
produced by the emitted radiations of unknown and
internal standard elements oppose each other through
galvanometer.
•P1 is used to balance opposing currents. Settings of P1 is
calibrated in terms of known solutions. P2 is used for
sensitivity adjustments.
Recording type flame photometer
 Recording type Flame Photometer
•Air is introduced around atomizer nozzle
through inlet F at a controllable pressure.
•Gas enters mixing chamber through M. The
sample B is drawn up the capillary tube C.
Large droplets pass through drain tube E.

•Baffles G ensure thorough mixing of gas, air

and samples.
•Light from flame is collected by reflector H.
The critical wavelength is isolated by the
interchangeable filter J.
 Clinical Flame Photometer
•The flame photometer is one of the most useful
instruments in clinical analyses. This is due to
the suitability of the flame photometer for
determining sodium (Na) , potassium (K) and
calcium (Ca), which are of immense importance
in the development of the living being and
indispensable to its physiological functions.
•In the clinical analysis of sodium and
potassium, the flame photometer gives, rapidly
and accurately, numerous differential data for
normal and pathological values.
 Clinical Flame Photometer
•The normal plasma sodium concentration is
about 140 mmol/l. A high plasma sodium
concentration of more than 145 mmol/l is
referred to as hypernatremia. This can occur
due to simple dehydration, excess sodium
intake, steroid therapy as well as in diabetic
insipidus.
•Hyponatremia, with plasma sodium
concentration less than 130 mmol/l, can occur
due to diuretic medication, kidney disease,
excessive sweating, congestive heart failure or
gastrointestinal disorder.
 Clinical Flame Photometer
•Potassium is the major intracellular action and
influences muscular activity, cardiac function
and nerve conduction process.
•Acute hyperkalaemia condition in which the
plasma potassium concentration exceeds 5.5
mmol/l is a medical emergency.
•In hypokalaemia, the potassium level will be
less than 3.5 mmol/l, which can occur due to
excessive loss in gastrointestinal secretions and
urine.
 Clinical Flame Photometer

•KLiNa Flame System of M/S Beckman is a

dedicated instrument for simultaneous analysis
of sodium and potassium.
•In this instrument, sample handling is
automatic, as the system has a turntable, which
will hold up to 20 samples in cups and an
automatic positive piston displacement dilutor
that dilutes the sample prior to entering the
spray chamber
Clinical Flame Photometer
Clinical Flame Photometer
Schematic diagram
 Clinical Flame Photometer
•For the precise and accurate determination of
sodium (Na) and potassium (K)
concentrations, use is made of the fact that
lithium normally not present in significant
concentrations in serum, exhibits about the
same flame emission characteristics as Na and
K.
•Lithium ions are added to the diluent used for
samples, standards and controls. The lithium
in the diluent is referred to as the internal
standard.
 Clinical Flame Photometer
•The diluted sample containing the fixed known
amount of lithium, in the form of a dissolved
salt, is nebulised and carried by the air supply
into the first of two compartments in the spray
chamber .
•Heavier droplets fall out of the stream on to
the chamber walls or separate from the stream
upon striking a partition in the chamber and
flow to a drain from the compartment.
 Clinical Flame Photometer
•Propane enters the first chamber to mix with the air
and sample stream, and carry it through a tubular
glass bridge into a second compartment; the aerosol
and propane mixture travels up from the chamber to
the burner head, where the mixture is burned.
Exhaust gases are vented to room air from a cover
located on top of the instrument.
•In order to provide internal standardisation, the
response of the sodium and potassium detector is a
ratio function of the response by the lithium detector.
Thus, any change in air-flow rate or fuel pressure that
may affect the sample would proportionately affect
the lithium detector.
 Clinical Flame Photometer
•The flame is monitored continuously by three
photomultiplier detectors. Each detector views
the flame through an optical filter that passes
only the wavelength band of interest to the
particular detector.
•The sodium detector therefore responds only
to wavelengths in a narrow band centred at 589
nm; the potassium detector responds only to
wavelengths in a narrow band centred at 766
nm; the lithium detector responds only to
wavelengths in a narrow band centred at 671
nm.
 Clinical Flame Photometer
•The specifications for the KLiNa Flame System
are +0.2 mmol/l for Potassium and Lithium,
and ±2.0 mmol/l for sodium.
•Potassium and lithium both show linearity to
20 mmol/l, while sodium is linear to 200. In
addition to the 0–20 scale, potassium may be
re-scaled to read out to 200 mmol/l for
convenient analysis of urine samples.