UNIT-2

Spectroscopic Techniques– I
 Outline
• The Electromagnetic Spectrum
• Classification of spectroscopic techniques
• Laws relating to absorption of radiation
• Block diagram of an absorption instrument
• Sources of absorption instrument
• Detectors
• Optical components for photometers
• Colorimeter/Photometer
• Infrared Spectroscopy
• Fourier Transform Infrared Spectroscopy
• UV – VIS spectrophotometer
 Electromagnetic spectrum

Figure: Various regions in the electromagnetic spectrum which are normally used
in spectroscopic work

Courtesy: A Handbook of Analytical Instruments by R.S.Khandpur,3rd Edition

 Electromagnetic spectrum

As wavelength increases frequency decreases.

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 Classification of
Spectroscopic Techniques
Classification of spectroscopic techniques
The different types of spectroscopy
include:
a. Based on the level of study
b. Based on the property of either
absorption or emission.
c. Based on the level of study i.e.
electronic or magnetic levels.
a) Based on the level of study- Here the
classification is done based on the study made at atomic or
molecular level. When light is allowed to pass through the
substance, changes in atomic level or molecular level is
observed.
1) Atomic spectroscopy: Here energy changes takes place
at atomic levels. The measurement is done to study the
atoms and their quantity. There are two types: atomic
absorption spectroscopy and atomic emission spectroscopy.
2) Molecular spectroscopy: Here the energy changes
occurring at the level of molecule are studied. The
characters like molecular absorption, emission and vibration
are studied. The examples include, colorimetry, UV-
spectroscopy, infrared, FTIR, fluorimetry etc.
B) Based on the property of either absorption or
emission.
Here the principle of absorption or emission of
electromagnetic radiation is taken into consideration.
1) Absorption spectroscopy: As the name suggests,
here there is absorption of light by the sample. The
extent of absorption and the wavelength of the
absorbed light is considered. The wavelength of light
absorbed tells the nature of the compound while
the intensity of absorbed light tells the concentration
• The examples of the spectroscopic methods
are colorimetry, UV-spectroscopy, infrared
spectroscopy, NMR spectroscopy, atomic
absorption spectroscopy.
2) Emission spectroscopy: Here the emitted light is
measured. Once the light impinge on the sample,
some of it is absorbed.
• This absorption of light leads to transition of electrons
from ground state to excited state.
• These excited electrons return back to ground state
by release of electromagnetic radiation (light) of
specific wavelength.
• The intensity gives the concentration while the
wavelength tells the nature of the compound.
• Examples of this method are fluorescence
spectroscopy, flame photometry.
C) Based on the level of study i.e. electronic or
magnetic levels.
Here the study is done based on electronic or magnetic
properties of the compound. Light is an electromagnetic
radiation. That is it has both electronic and magnetic
properties.
• Electronic spectroscopy: When a compound is estimated
without the magnetic field we call it electronic spectroscopy.
In this method the substance under test is exposed to light
without the influence of magnetic field.
• Examples of this method are colorimetry, UV visible
spectroscopy, IR, fluorimetry etc.
• Magnetic spectroscopy: Here the substance is exposed to
electromagnetic radiation in presence of external magnetic
field.
• Examples include Nuclear magnetic resonance spectroscopy
(NMR), Electron spin resonance spectroscopy (ESR).
 Laws relating to
absorption of radiation
Laws relating to absorption of radiation

• 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.
• Laws relating to absorption of radiation:
- Lambert’s Law
- Beer’s Law
- The Beer-Lambert Law
 Lambert’s Law
• Lambert’s Law states that each layer of equal thickness of
an absorbing medium absorbs an equal fraction of the
radiant energy that traverses it.
• Let us suppose, I0 is the incident radiant energy and I is the
energy which is transmitted.
• The ratio of the radiant power transmitted by a sample to
the radiant power incident on the sample is known as the
transmittance.
• Lambert’s Law is expressed as:
Transmittance T = I/ I0
• It is usually expressed as a percentage
% Transmittance = I/I0 × 100
• The logarithm of the reciprocal of the transmittance is
known as absorbance.
• Absorbance = log10 (1/T) = log10 (I0/I)
 Beer’s Law
• This law states that the absorption of light is
directly proportional to both the concentration of
the absorbing medium and the thickness of the
medium in the light path.
 The Beer-Lambert Law
• A combination of the two laws, known jointly as the Beer-Lambert
Law defines the relationship between absorbance (A) and
transmittance (T).
• It states that the concentration of a substance in solution is
directly proportional to the ‘absorbance’, A, of the solution.
Absorbance=𝑨=𝒄𝝐𝒃
where: 𝐴 = absorbance
𝑐 = concentration
𝜖 = molar absorptivity (how much light is absorbed by 1
mole of a molecule)
𝑏 = length of the path that the light travels
• ε is a function of wavelength. So, the Beer-Lambert Law is true
only for light of a single wavelength or monochromatic light.
• Absorptivity is constant, depending upon the
wavelength of the radiation and nature of the
absorbing material.
• Absorbance is related to percentage
transmittance T by the expression:
A = log10 (I0/I) = log10 (100/T) = εbc
 Block diagram of an
absorption instrument
 Block diagram of an absorption
instrument

Figure: Various components of an absorption instrument

Sources of absorption
instrument
 Radiation Sources
• The function of the radiation source is to
provide sufficient intensity of light suitable
for making a measurement.
• The most common and convenient source of
light is the tungsten lamp.
• Lamps convert electrical energy into
radiation.
• Different designs and materials are needed
to produce light in different parts of the
electromagnetic spectrum.
• Several types of lamps that are useful in spectroscopy
are as follows:
Blackbody sources:
• A hot material, such as an electrically heated filament
in a light bulb, emits a band of spectrum of light.
• The spectrum is approximated by Planck’s radiation
law for blackbody radiators:
 
 v 
3
1 
B  2 h 2  
 c  exp h  1
 kT 

Where h is Planck’s constant,  is frequency, c is the

speed of light, k is the Boltzmann constant, and T is
temperature in K.
• The most common incandescent lamps and their
wavelength ranges are:
- Tungsten filament lamps : 350 nm–2.5 mm
- Glowbar : 1–40 mm
- Nernst glower : 400 nm–20 mm
Figure: A tungsten lamp

• Tungsten lamps are used in visible and near infrared

(NIR) absorption spectroscopy, glowbar, and Nernst
glower are used for infrared spectroscopy.
• A major portion of the energy emitted by a tungsten
lamp is in the visible region and only about 15–20% is
in the infrared region.
• For work in the UV region, a hydrogen or deuterium-
discharge lamp is used.

Courtesy: A Handbook of Analytical Instruments by R.S.Khandpur,3rd Edition

• The radiation from the discharge lamps is
concentrated into narrow wavelength regions of
emission lines.
• Practically, there is no emission beyond 400 nm
in these lamps.
• Nernst filaments are operated at lower
temperatures and radiates sufficient energy.
• The curves in figure show the energy distribution
for a tungsten filament at three different
temperatures.
• Such radiation is known as ‘black body radiation’.
• In the figure, we can observe that how the
emitted energy increases with temperature and
how the wavelength of maximum energy shifts to
shorter wavelengths.

Figure: Tungsten filament radiation characteristics

Courtesy: A Handbook of Analytical Instruments by R.S.Khandpur,3rd Edition

The tungsten-halogen lamp:
• The introduction of the tungsten-halogen light
source has a higher intensity output than the
normal tungsten lamp in the region of 320–
380nm and are used in colorimetry,
spectrophotometry.
• The tungsten filament evaporates by releasing
particles.
• The evaporated tungsten combines with halogen
gas within the glass envelope to create tungsten
– halogen molecule.
• The tungsten – halogen molecule then migrates
back to the filament, eliminating blackening of
the glass envelope.

• The tungsten is re-deposited and recycled onto

the filament, strengthening the filament,
extending the life of the lamp and the halogen
gas is then free to start the cycle again.
Figure: The tungsten-halogen lamp
Deuterium tungsten-halogen light source:
• A combination of deuterium tungsten-halogen light
source combines the continuous spectrum of a
deuterium UV light source and a tungsten-halogen
VIS/Shortwave NIR light source in a single optical
path.
• The combined light source produces a powerful, stable
output from ~200–1,100 nm.
 Discharge lamp
• Discharge lamps, such as neon, pass an electric current
through a rare gas or metal vapour to produce light.
• The electrons collide with gas atoms, exciting them to higher
energy levels which then decay to lower levels by emitting
light.
• Common discharge lamps and their wavelength ranges are
given below:
- Hydrogen or deuterium: 160–360 nm
- Mercury: 253.7 nm, and weaker lines in the near UV and
visible
- Ne, Ar, Kr, Xe discharge lamps: many sharp lines throughout
the near UV to near-lR
- Xenon arc: 300 – 13 nm
• Deuterium lamps are the UV
source in UV-Vis absorption
spectrophotometers.
• For fluorescent work, an intense
beam of UV light is required. This
requirement is met by a Xenon arc
or a mercury vapour lamp.
• Mercury lamps are usually run
direct from the AC power line via a
series ballast choke (fast warm up
to the lamp to obtain operating
temperature is).
Figure: A deuterium lamp of the
type used in spectrophotometers

Courtesy: A Handbook of Analytical Instruments by R.S.Khandpur,3rd Edition

• A deuterium arc lamp provides
emission of high intensity and
adequate continuity in the range of
190–380 nm.
• A quartz or silica envelope is
necessary not only to provide heat
shield, but also to transmit the
shorter wavelengths of the UV
radiation.
• Figure shows the energy output as
a function of wavelength in case of
deuterium arc lamp and tungsten-
halogen lamp.
Figure: Energy output as a
function of wavelength for
deuterium arc lamp and
tungsten halogen lamp

Courtesy: A Handbook of Analytical Instruments by R.S.Khandpur,3rd Edition

 Lasers
• The term laser has been coined by taking the first letters of
the expression ‘Light amplification by simulated emission of
radiation’.
• The laser beam has spatial and temporal coherence, and is
monochromatic (pure wavelength).
• The beam is highly directional and exhibits high density
energy which can be finely focused.
• The advantages include higher intensity, monochromaticity,
low beam divergence, the availability of short and
ultrashort pulses for studies of transient phenomena, and
coherence (well-defined phase).
• Lasers have been used in many spectroscopic applications
such as Raman Spectroscopy, UV/visible
spectrophotometry.
 Light Emitting Diodes (LEDs)
• The development of LEDs has resulted in the appearance of a new
optical light source in analytical instrumentation.
• The first LED was developed in 1962 based on GaAsP layers,
which emitted red light.
• Significant advances in III-V nitride manufacturing processes have
resulted in high power commercially available LEDs in the region
of 247–1,550 nm, covering UV, visible and near infrared regions.
• The advantages include increased lifetime, low cost, reduced
power consumption, higher brightness, rugged construction,
flexible configuration, enhanced spectral purity, small size, and
breadth of spectral range.
• The most popular type of tri-colour LED has a red and a green
LED combined in one package with three leads.
• When both the red and green LEDs are turned on, the LED
appears to be yellow.
• The centre lead (K) is the
common cathode for both
LEDs, the outer leads (A1 and
A2) are the anodes to the LEDs
allowing each one to give
separately, or both together to
give the third colour.
• The use of bi-/tri- colour LEDs
can provide a compact rugged
multi-wavelength photometer
source that can facilitate multi-
Figure: Schematic of Tri-colour LED
component analysis.

Courtesy: A Handbook of Analytical Instruments by R.S.Khandpur,3rd Edition

Detectors
 Detectors
• Photosensitive Detectors
- Photovoltaic or barrier layer cells
- Photo-emissive cells
. High vacuum photo-emissive cells
. Gas-filled photo-emissive cells
. Photomultiplier tubes
• Silicon diode detectors
• Photodiode arrays (PDA)
 Photosensitive Detectors
• After isolation of radiation of a particular
wavelength in a filter or a monochromator, it is
essential to have a quantitative measure of
their intensities.
• This is done by causing the radiation to fall on a
photosensitive element, in which the light
energy is converted into electrical energy.
• The electric current produced by this element
can be measured with a sensitive galvanometer
directly or after suitable amplification.
• There are two types of photoelectric cells:
photovoltaic cells and photo-emissive cells.
 Photovoltaic or barrier layer
cells
• Photovoltaic or barrier-layer cells
usually consist of a semiconducting
substance, which is generally selenium
deposited on a metal base which may
be iron and which acts as one of the
electrodes.
• The semiconducting substance is
covered with a thin layer of silver or
gold deposited by cathodic deposition in
vacuum. This layer acts as a collecting
electrode.
• Figure shows the construction of the
barrier-layer cell. Figure: Construction of a barrier layer cell
• When radiant energy falls upon the
semiconductor surface, it excites the
electrons at the silver-selenium
interface.
• The electrons are thus released and
collected at the collector electrode.
• Photovoltaic cells are very
robust in construction and no
need of external electrical
supply to work.
• The typical photocurrents
produced by these cells are as
high as 120 mA/lumen.
• At constant temperature, the
current set up in the cell
usually shows a linear
relationship with the incident
light intensity.
• Selenium cells are sensitive to Figure: Spectral response of a selenium
almost the entire range of photocell and the human eye
wavelengths of the spectrum
 Photo-emissive cells
• Photo-emissive cells are of three types:
(a) high vacuum photocells,
(b) gas-filled photocells and
(c) photomultiplier tubes.
• All of these types differ from selenium cells, in that
they require an external power supply to provide a
sufficient potential difference between the electrodes
to facilitate the flow of electrons generated at the
photosensitive cathode surface.
• Also, amplifier circuits are invariably employed for the
amplification of this current.
 High vacuum photo-emissive cells
• The vacuum photocell consists of two electrodes, cathode
having a photosensitive layer of metallic caesium deposited
on a base of silver oxide and the anode is either an axially
centered wire or a rectangular wire that frames the
cathode.
• The construction of the anode is such that no shadow falls
on the cathode.
• The two electrodes are sealed within an evacuated glass
envelope.
• When a beam of light falls on the surface of the cathode,
electrons are released from it, which are drawn towards the
anode which is maintained at a certain positive potential.
• This gives rise to photocurrent, which can be measured in
the external circuit.
• The spectral response of a photo-emissive tube depends on
the nature of the substance coating the cathode.

Figure: Typical circuit configuration

employed with photo-emissive tubes
Courtesy: A Handbook of Analytical Instruments by R.S.Khandpur,3rd Edition
• Figure shows current-voltage characteristics of vacuum
photo-emissive tube at different levels of light flux.
• They show that as the voltage is increases, the point is
reached where all the photoelectrons are swept to the anode
as soon as they are released and result in saturation
photocurrent.

Figure: Current-voltage characteristics of

vacuum
Courtesy: A Handbook of Analytical Instruments by R.S.Khandpur,3rd Edition
Gas-filled photo-emissive cells
• This type of cell contains small quantities of
inert gas like argon, whose molecules can be
ionised when the electrons present in the cell
possess sufficient energy.
• The presence of small quantities of this gas
prevents the phenomenon of saturation
current, when higher potential differences are
applied between the cathode and anode.
• Due to repeated collisions of electrons in the
gas-filled tubes, the photoelectric current
produced is greater even at low potentials.
 Photomultiplier tubes
• Photomultiplier tubes (PMTs) convert photons to an electrical
signal.
• They have a high internal gain and are sensitive detectors for low-
intensity applications such as fluorescence spectroscopy.
• Photomultiplier tubes are used as detectors when it is required to
detect very weak light intensities.
• The tube consists of a photosensitive cathode and has multiple
cascade stages of electron amplification; in order to achieve a
large amplification of primary photocurrent within the envelope of
the phototube itself.
• When a photon of sufficient energy strikes the photocathode, it
ejects a photoelectron due to the photoelectric effect.
• The photocathode material is usually a mixture of alkali metals,
which make the PMT sensitive to photons throughout the visible
region of the electromagnetic spectrum.
• The photocathode is at a high negative voltage,
typically −500 to −1,500 V.
• The photoelectron is accelerated towards a series of
additional electrodes called dynodes.
• These electrodes are each maintained at successively
less negative potentials.
• Additional electrons are generated at each dynode.
There may be 9–16 dynodes.
• This cascading effect creates 105 to 107 electrons for
each photoelectron that is ejected from the
photocathode.
• The amplification depends on the number of dynodes
and the accelerating voltage.
• This amplified electrical signal is collected at an anode
at ground potential, which can be measured.
• The sensitivity of the
PMT can be varied by
regulating the voltage of
the first amplifying
stage.
• Because of the relatively
small potential
difference between the
two electrodes, the
response is linear.
Figure: Schematic of a photomultiplier tube
• The output of the PMT is
limited to an anode
current of a few milli
amperes.
• A direct current (DC) power supply is required to
operate a photomultiplier, the stability of which must
be at least one order of magnitude better than the
desired precision of measurement.
• The devices are sensitive to electromagnetic
interference, and they are also more costly than other
photoelectric sensors.
• Photomultipliers are not uniformly sensitive over the
whole spectrum.
• In the case of spectrophotometers, the
photomultipliers normally supplied cover the range of
185–650 nm.
• For measurements at longer wave lengths, special
red-sensitive tubes are offered.
 Silicon diode detectors
• The photomultiplier which is large and expensive and requires a source of
stabilised high voltage, can be replaced by a photodiode.
• This diode is useable within a spectral range of 400–1,050 nm, in a
number of instruments (spectrophotometers, flame photometers).
• When a photon strikes a semiconductor, it can promote an electron from
the valence band (filled orbital) to the conduction band (unfilled orbital)
creating an electron (−) – hole (+) pair.
• The concentration of these electron-hole pairs is dependent on the amount
of light striking the semiconductor, making the semiconductor suitable as
an optical detector.

Figure: Schematic of a semiconductor detector

• There are two ways to monitor the
S.No. Detector λ(μ)
concentration of electron-hole pairs. type
• In photodiodes, a voltage bias is 1. Si 0.2–1.1
present and the concentration of
2. Ge 0.4-1.8
light-induced electron-hole pairs
3. InAs 1.0-3.8
determines the current through
4. InSb 1.0–7.0
semiconductor.
• Photovoltaic detectors contain a p-n 5. InSb (77K) 1.0–5.6
junction that causes the electron-hole 6. HgCdTe 1.0–25.0
pairs to separate to produce a (77K)
voltage that can be measured.
•They cover a spectral range from Table: The wavelength
185 to 850 nm. range for different solid-
state detectors
• The figure shows spectral response of silicon
diode detectors.
• The devices being solid state are mechanically
robust and consume much less power.

Figure: Spectral response of silicon diode detectors

 Photodiode arrays (PDA)
• Diode arrays are assemblies of
individual detector elements in
linear or matrix form, which in
a spectrophotometer can be
mounted so that the complete
spectrum is focused on to an
array of appropriate size.
• The arrangement does not
require any wavelength Figure: Schematic of a photodiode array
selection mechanism and the
output is instantaneously
available.
• However, resolution in diode
arrays is limited by the physical
size of individual detector
elements.
• A photodiode array consisting of discrete photodiodes is
available on an integrated circuit (IC) chip.
• Array detectors are especially useful for recording the full
uv-vis absorption spectra of samples that are rapidly
passing through a sample flow cell, such as in HPLC
detectors.
• PDAs are available with 512, 1,024, or 2,048 elements with
typical dimensions of 25 mm wide and 1–2 mm high.
• Light creates electron-hole pairs and the electrons migrate
to the nearest PIN junction.
• After a fixed integration time the charge at each element is
read with solid-state circuitry to generate the detector
response as a function of linear distance along the array.
• The detector consists of two silicon integrated circuits, each
containing 211 photosensitive diodes and 211 storage
capacitors.
Optical components for
photometers
 Optical Components
• Several different types of optical components are used in
the construction of analytical instruments based on the
radiation absorption principle.
• They could be windows, mirrors and simple condensers.
• The following factors need to be considered while selecting
optical components:
- From 180 to 210 nm fused silica can be used.
- Below 300 nm, quartz or fused silica is utilised, the limit
for quartz is 210 nm.
- From 300 to 350 nm, special corex glass can be used
- Ordinary silicate glasses are satisfactory from 350 to
3,000 nm.
• Reflections from glass surfaces are reduced by coating
these with magnesium fluoride.
• Silica or synthetic quartz coating is used which is hard
and chemically resistant.
• The use of silica-coated aluminium mirrors ensures
long mirror life with enhanced reflectance in the UV
region and minimum deterioration of stray-light
performance.
 Table provides data on materials that are used for
optical components in different parts of the
electromagnetic spectrum.

S.No Region of Mirrors Lenses Windows

the
spectrum
1. X-ray –– –– Beryllium
2. Ultraviolet Aluminium Fused Silica Fused silica,
(Synthetic sapphire
Quartz),
Sapphire

3. Visible Aluminium Glass, sapphire Glass, sapphire

4. Near infrared Gold Glass sapphire Glass sapphire

5. Infrared Copper, CaF2, ZnSe NaCl, BaF2, CaF2,

gold SnZe

Courtesy: A Handbook of Analytical Instruments by R.S.Khandpur,3rd Edition

• In order to reduce the beam size or render the
beam parallel, condensers are used.
• These condensers operate as simple microscopes.
• To minimise light losses, lenses are sometimes
replaced by front-surfaced mirrors to focus or
collimate light beam in absorption instruments.
• Mirrors are aluminised on their front surfaces.
With the use of mirrors, chromatic aberrations
(deviations) and other imperfections of the lenses
are minimised.
• Beam splitters are used in double-beam
instruments. These are made by giving a suitable
multilayer coating on an optical flat.
• The two beams must retain the spectral
properties of the incident beam.
• Half-silvered mirrors are often used for splitting the
beam.
• However, they absorb some of the light in the thin
metallic coating.
• Beam splitting can also be achieved by using a
prismatic mirror or stack of thin horizontal glass
plates, silvered on their edges and alternatively
oriented to the incident beam.
• Fibre Optics: Optical fibres are now being
extensively used in spectroscopic applications to
transmit light for quantitative measurements.
• The fibres used in most of the analysers have core
diameters of 200–600 microns.
• They come in two types based upon their hydroxyl
content. (Low-OH fibres and High-OH fibres).
Colorimeter/Photometer
 Introduction
• A colorimetric method in its simplest form uses only
the human eye as a measuring instrument.
• This involves comparison by visual means of the
colour of an unknown solution, with the colour
produced by a single standard or a series of
standards.
• A colorimetric determination is one that involves
visual measurement of color; and a method
employing photoelectric measurement is referred as a
photometric or spectrophotometric method.
• Usually any method involving measurement of color in
the visual region of the electromagnetic spectrum
(400–700 mm) is referred to as the colorimetric
method.
• In a colorimeter, the sample is normally a liquid.
• The sample compartment of a colorimeter is
provided with a holder to contain the cuvette, in
which the liquid is examined.
• Usually this holder is mounted on a slide with
positions for at least two cuvettes, so that sample
and reference cuvettes are measured first and a
shutter is moved into or out of the light beam
until the micro-ammeter gives a full-scale
deflection (100% T-scale reading).
• The sample is then moved into the beam and the
light passing through it is measured as a
percentage to the reference value.

samplereading
sampleconcerntration  s tan dardconcentration 
Re ferencereading
 Types of colorimeter/Photometer

• Single-beam Filter Photometers

• Double-beam Filter Photometer
• Probe Type photometer
• Miniature Fibre Optic Spectrometer
• Multi-channel Photometer
• Pocket Colorimeter
• Process Photometers
 Single-beam Filter Photometers

Figure: Basic components of a filter photometer

Courtesy: A Handbook of Analytical Instruments by R.S.Khandpur,3rd Edition

 Working of Single-beam Filter Photometers
• The source of light is a tungsten filament lamp, which
is held in a reflector and throws light on the sample
holder through a filter.
• The filter may be either of absorption or interference
type.
• The sample holder is a cuvette with parallel walls or
may be a test tube.
• The light, after passing through the sample holder,
falls on the surface of the photocell.
• The output of the cell is measured on a micro-
ammeter.
• The lamps must be energised from a highly stabilised
DC source, or by the output of a constant voltage
transformer.
Double-beam Filter Photometer
• In double-beam filter photometers, two photocells are
normally employed.
• The two photocells are connected to two potentiometers R1
and R2.
• Each of the two potentiometers is of low resistance and is
wound linearly.
• Light from the source lamp is made to pass through the
filter and is then divided into two parts, one part passing
through the solution in the cuvette before falling on the
measuring photocell, and the other part passing directly on
to the reference photocell.
• The galvanometer G receives opposing currents through it.
• The potentiometer R1 is measured in transmittance and
absorbance units.
 Figure: Schematic of a double-beam filter photometer

Courtesy: A Handbook of Analytical Instruments by R.S.Khandpur,3rd Edition

• With the lamp off, the galvanometer zero is
adjusted mechanically.
• The potentiometer R1 is set to T = 1 or A = 0.
• Then with the lamp on, the blank solution is
placed in the light path of the measuring cell.
• Potentiometer R2 is adjusted until galvanometer
G reads zero.
• The solution to be analysed is then substituted
for the blank and R1 is adjusted until the
current through the galvanometer is zero, the
setting of R1 remaining unchanged.
• The absorbance or transmittance can then be
read directly on the scale of potentiometer R1.
• In two-cell photometers, the errors resulting from
the fluctuations of the lamp intensity are
minimised.
• The scale of potentiometer R1 (transmittance
scale) can be made much larger in size than the
scale of the metre in single-cell instruments.
• They directly display the absorbance values on
direct-reading digital display.
 Probe Type photometer
• Dipping probe-type photometers use fibre optics to transport
light.
• The light from a tungsten lamp travels down a fibre optic
cable which is dipped into the solution of interest (sample).
• The light then passes through the solution and a mirror
reflects the light back to a return fibre optics cable.
• The cell path length is two times the distance between the
ends of the optical cable and the mirror.
• Interference filters are provided to select a wavelength.
• The reflected light is then passed through a photodiode with
an amplifier and an electronic chopper which is synchronised
with the lamp.
• This results in the detector not responding to extraneous
light.
Schematic diagram of a probe-type photometer

Figure: Schematic diagram of a probe-type photometer

Courtesy: A Handbook of Analytical Instruments by R.S.Khandpur,3rd Edition
Miniature Fibre Optic Spectrometer
• Miniature fibre optic spectrometer, the S2000 is
shown below:
• The total system comprises five basic elements:
the S2000 Miniature Fibre Optic Spectrometer, an
A/D converter, operating software, a light or
excitation source, and sampling optics.

Figure: Absorbance/ Transmission Setup

Courtesy: A Handbook of Analytical Instruments by R.S.Khandpur,3rd Edition

• The light or excitation source sends light through an optical
fibre to the sample.
• The light interacts with the sample. Then the light is
collected and transmitted through another optical fibre to
the spectrometer.
• The spectrometer measures the amount of light and the
A/D converter transforms the analog data collected by the
spectrometer into digital information that is passed to the
software, providing the user with application-specific
information.
• The software has the ability to perform spectroscopic
measurements such as absorbance, reflectance and
emission; control all system parameters; collect data from
up to eight spectrometer channels simultaneously in real
time, display the results in a single spectral window; and
perform reference monitoring and time acquisition
experiments.
• The optical system has two identical fibres in a bifurcated
assembly.
• A plano-convex lens shapes the light coming out of the
illumination fibre.
• The light is transmitted through the sample, hits the mirror,
reflects off the mirror, and interacts with the sample again
before being transmitted back through the probe via the
read fibre.
• Because the light travels through the sampling region twice,
the optical path length is actually twice the length of the
sample aperture.
• The transmission cell is used to measure absorbance of the
fluid that fills the sample compartment between the fibres
and the mirror.
• The silica-core and silica-clad optical fibres are optimised for
the UV-VIS (200–750 nm) or VISNIR (450–1,000 nm)
range.
 Multi-channel Photometer
• An increasing number of chemical analysis are
carried out in the laboratories of industries and
hospitals by a photometer.
• One of the limitations for rapid analyses is the
speed at which the samples can be transferred in
the light path.
• In a multi-channel photometer, instead of
introducing one sample at a time into a single
light path, a batch of samples is introduced and
measurements carried out simultaneously, using
a multiplicity of fibre optic light paths and
detectors, and then scanned electronically
instead of mechanically.
 Figure: Schematic of a multi-channel photometer

Courtesy: A Handbook of Analytical Instruments by R.S.Khandpur,3rd Edition

• The 24 sample cuvettes are arranged in a rack in a
three key eight matrix.
• The 25th channel serves as a reference beam and
eliminates possible source and detector drifts.
• The light source is a 50 W tungsten-halogen lamp,
driven from a precisely controlled voltage source.
• The light is chopped by means of a mechanical
rotating chopper.
• A lens focuses the light on the end of a bundle of fibre
optic elements.
• The output of the detectors is amplified and displayed
on digital volt-metre.
• The whole operation is synchronised with digital logic
circuits.
 Pocket Colorimeter
• Pocket Colorimeter is lightweight, battery
operated, waterproof and ideal for field work.
• It has accuracy comparable to a lab instrument,
yet it remains simple to use.
• It operates on fixed wavelengths 420, 450,
476, 500, 550, 580, 600, 650 nm with
bandwidth of 15 nm.
• The use of multi-LED photometers allows a
wide range of the electromagnetic spectrum to
be covered simultaneously or individually
without manually changing the LEDs.
• Hauser and few more authors employed a fibre
optic coupler to guide the light from up to 7 LEDs
into a single measuring cell.
• This photometer could be configured to detect Al,
Cu, NH3, Cu, Ca, chromium, phosphate and
nitrite using colorimetric methods.
• A coupler was used to merge the light from one
of the seven input channels into two output
fibres.
• One was brought to the measuring photodiode
and the other was brought to the reference
photodiode
• The photometer features a data logging function
that allows users to store and recall the ten most
recent data points, eliminating the need to
manually record data.

Figure: The circuit diagram for the multi-LED photometer

Courtesy: A Handbook of Analytical Instruments by R.S.Khandpur,3rd Edition

 Process Photometers
• If we need to monitor an analyte’s concentration
over time, it may not be possible to physically
remove samples for analysis.
• Examples: when monitoring industrial production
lines or waste lines, when monitoring a patient’s
blood, or when monitoring environmental
systems. With a fibre-optic probe, we can analyse
samples.
• The fibre optic probe with a membrane containing a reagent that
reacts with the analyte. When the analyte diffuses across the
membrane, it reacts with the reagent, producing a product that
absorbs UV or visible radiation.
• The non-absorbed radiation from the source is reflected or
scattered back to the detector. Fibre optic probes that show
chemical selectivity are called optrodes.

Figure: Example of a fibre-optic probe

Courtesy: A Handbook of Analytical Instruments by R.S.Khandpur,3rd Edition

 Block diagram of a single probe
instrument

Figure: Optical diagram of a single channel uv-vis and NIR process

photometer

Courtesy: A Handbook of Analytical Instruments by R.S.Khandpur,3rd Edition

• Light is sent to the fire optic probe and returned to the analyser.
• Beam splitters divide the light and a portion passes through
narrow-band wavelength filters into each detector.
• Up to six detectors, one or more of which are references may be
used.
• Any combination of NIR (In GaAs) and UV-Vis (Si) detectors can
be placed in any position, providing flexibility in operation.
• Voltage from each detector is logarithmically converted, and then
the signals from each peak and its reference are subtracted,
producing a voltage equivalent to optical absorbance units (Au).
• A microprocessor converts the signals into chemical units for
display and transmits them as 4–20 mA signals to the process
computer.
• The instrument uses reference voltage (i.e. the signal strength at
the reference wavelength) to identify fibre breakage, probe
fouling or intermittent cloudiness due to filter break through.
• It additionally uses an internal circuit to indicate if the lamp is
about to burn out or if it has burnt out.
Infrared Spectroscopy
 Infrared Spectroscopy- Introduction
• Infrared (IR) spectrophotometer is ideally suited for carrying out
qualitative and quantitative analysis, particularly of organic
compounds.
• IR region extends from 0.8 to 200 m in the electromagnetic
spectrum. But most of the commercial instruments are available in
the region from 0.8 to 50 m.
• The position of absorption bands in the IR spectrum is expressed
both in wavelength as well as in wave numbers.
• The wavelength λ is generally measured in microns (μ).
• The wavenumber (u) is the number of wavelengths per centimetre
and is given by
ν (in cm−1) = 104/λ (in μ)
• IR spectrophotometers using prisms produce spectra which are
spread linearly with wavelength, whereas instruments fitted with
gratings generally deliver spectra spread linearly with wave
number.
Various regions of the IR range of the
spectrum

Region Wavelength Wavenumber

range (λ m) range (cm−1)
Near 0.78–2.5 12800–4000
Middle 2.5–50 4000–200
Far 50–1000 200–10
Basic Components of IR Spectrophotometers
• Spectrophotometers for the IR range are
composed of the same basic elements, as
instruments in the visible and ultraviolet range,
namely:
- a source of radiation
- a monochromator for dispersing the radiation
- a detector which registers the residual intensity
after selective absorption by the sample
Block diagram of IR Spectrophotometers
 Radiation Sources
• The source of radiation in IR spectrophotometers
is ideally a black body radiator.
• The energy emitted by a black body radiator
varies with wavelength and with temperature.
• In particular, increasing the temperature of the
source raises the energy of emission enormously
in the short wavelength region, but has relatively
small effect at long wavelengths.
• The optimum IR source is an inert solid heated
electrically to temperatures between 1500 and
2000 K. The maximum radiant intensity at these
temperatures occurs at 1.7–2 m (5000–6000
cm−1).
 Types of IR sources
• There are three common practical IR sources: the
Globar rod, the Nernst filament and the Nichrome
wire.

Figure: Typical spectral characteristics of some IR sources

(a) Globar, (b) Nernst glower, (c)Tungsten glower
 Globar
• The Globar is a silicon carbide rod, which has a positive
temperature coefficient of resistance.
• The rod is about 5 cm in length and 0.5 cm in diameter. It is
electrically heated and run at a current to produce a temperature
of about 1300 K.
• The heat dissipation at the ends of the rod is high,
as they are the coolest parts. This sometimes leads to
arcing and burnout.
• Therefore, water cooling of electrical parts is required to prevent
arcing.
• The Globar finds applications for work at wavelengths
longer than 15 m (650 cm−1), because its radiant
energy output decreases less rapidly.
 Nernst filament
• The Nernst filament is a small rod composed of
fused rare earth oxides of zirconium and yttrium.
• The filament is of cylindrical shape having a
diameter of about 1–2 mm and a length of 20–30
mm.
• Platinum leads are sealed to the ends of the
cylinder to allow passage of current through it.
• The filament is heated to temperatures between
1500 and 2000°C and produces maximum
radiation at about 7100 cm−1.
• The device has a negative temperature coefficient
of electrical resistance and is, therefore, operated
in series with ballast resistance in constant voltage
circuit.
• It must be externally heated to a dull-red hot,
because it is non-conducting when cold and has a
tendency to crack or separate from its connections
on cooling.
• Therefore, it should be run continuously whenever
possible.
 Nichrome strip
• The nichrome strip, though, gives less energy than the
Globar or Nernst, and is extremely simple and reliable in
operation.
• In construction, it could be a tightly wound spiral of
nichrome wire, heated by passage of current.
• The temperature is about 800–900°C. Some IR
spectrophotometers employ a ceramic rod source, in which
is embedded a platinum-rhodium filament producing
temperature up to 1200°C.
 Monochromators
• Light from the entrance slit is rendered parallel after
reflection from a collimating mirror and falls on the
dispersing element.
• The dispersed light is subsequently focused on to the
exit slit of the monochromator and passes into the
detector section.
• The sample is usually placed at the focus of the beam,
just before the entrance slit to the monochromator.
• IR monochromators generally employ several mirrors
for reflecting and focusing the beam of radiation in
preference over lenses to avoid problems with
chromatic aberrations.
• Both prisms and gratings are used for dispersing
IR radiation.
• As a general rule, instruments operating below
25–40 m have prism monochromators, whereas
reflection gratings are utilised above 40 m,
because transparent materials are not easily
available in that range.
• Materials for prism construction, which are found
most suitable in the IR region.

Material Optimum range

Glass (Si02) 300 mm to 2 m (5000 cm−1)

Quartz 800 mm to 3 m (3300 cm−1)

Lithium fluoride 600 mm to 6.0 m (1670

cm−1)
Calcium fluoride 200 mm to 9.0m (1100
cm−1)
Sodium chloride 200 mm to 14.5 m (625
cm−1)
Potassium bromide 10 to 25 m (400 cm−1)

Caesium iodide 10 to 38 m (260 cm−1)

• The use of gratings as dispersing elements in the
monochromators offers a number of advantages for the IR
region.
• Better resolution is possible, because there is less loss of
radiant energy than in a prism system.
• This facilitates employing of narrower slits. Also, gratings offer
nearly linear dispersion.
• With these advantages, gratings have almost replaced prisms.
• The standard grating used in IR spectrophotometry is an
echelon reflection grating.
• They are usually constructed from glass or plastic that is
coated with aluminium.
• Grating instruments incorporate a sine-bar mechanism to drive
the grating mount, when a wavelength readout is desired and
a cosecant-bar drive when wave numbers are desired.
• One disadvantage of using gratings is that, they disperse
radiant energy into more than one order.
 Entrance and Exit Slits
• Most of the monochromators use a specially shaped cam to
control the slit width, as he wavelength is scanned.
• Besides this, a servo system can also be employed to
achieve the same purpose of the slit servo system, which
maintains the output of an optical monochromator at a
constant level, as the wavelength is scanned.
• The slit width control has the same importance as it has in
ultraviolet and visible spectrophotometers, and a
compromise of different factors is to be made while
selecting the slit width to be used.
• Narrow slits produce smaller bandwidth which consequently
results in better spectra definition, whereas wider slits
permit larger amount of radiant energy reaching the
detector and consequently greater photometric accuracy
 Mirrors
• As the materials that are used for lenses are not
transparent to IR radiation over the entire
wavelength range, lenses are generally not
preferred.
• Front surfaced mirrors are usually used in the IR
instruments, of which plane, spherical, parabolic
and toroidal types are the most common.
• Although highly reflecting aluminium-coated
mirrors with a protective coating are usually
employed
 Detectors
• Detectors used in IR spectrometers usually convert the thermal
radiant energy into electrical energy, which can subsequently be
plotted on a chart recorder.
• Selection of a specific detector depends on the waveband of
interest, the sensitivity required and the cost constraints.
• The detectors used may be divided into thermal and quantum
types.
• The quantum detectors are useful in the near IR region, whereas
thermal detectors can be used very much beyond this range.
• The main features of thermal-type detectors include responsively
with little dependence on wavelength and operation at room
temperature.
• However, the response speed and detectivity are lower than the
quantum type. The commonly used thermal detectors are
thermopiles, bolometers, pneumatic detectors and pyroelectric
detectors.
 Quantum-type detectors
• Quantum-type detectors feature high detectivity and fast response speed.
• Responsitivity is wavelength dependent and except for detectors in the
near IR range, cooling is normally used with these detectors.
• Quantum-type detectors are classified into intrinsic types and extrinsic
types.
• The wavelength limits of intrinsic-type detectors are determined by their
inherent energy gap, and responsitivity drops drastically when the
wavelength limit is exceeded.
• Typical examples of this type of detectors are photoconductive and
photovoltaic detectors such as Hg CdTe or PbSnTe.
• Extrinsic-types of detectors are photoconductive detectors whose
wavelength limits are determined by the level of impurities doped in high
concentrations to the Ge or Si semiconductors.
• The major difference between intrinsic-type detectors and extrinsic-type
detectors is the operating temperature.
• Extrinsic-type detectors must be cooled down to the temperature of liquid
helium.
• Quantum-type detector
- Photoconductive cells
- Solid-State Photo detectors
• Thermal detectors
These detectors depend on their response on the
heating effect of the radiation.
• Thermocouples, bolometers, pneumatic and
pyroelectric detectors are the three commonly
used thermal detectors
 Photoconductive Cell
• Light striking the surface of a material can provide
sufficient energy to cause electrons within the
material to break away from their atoms. Thus, free
electrons and holes (charge carriers) are created
within the material, and consequently its resistance is
reduced. This is known as the Photoconductive effect.
• Light-sensitive material is arranged in the form of a long
strip zigzagged across a disc-shaped base.
• The connecting terminals are fitted to the conducting
material on each side of the strip; they are not at the ends
of the strip.
• Thus, the light sensitive material is actually a short, wide
strip between the two conductors.
• Cadmium sulfide (CdS) and cadmium selenide (CdSe) are
the two materials normally used in photoconductive cell
manufacture.
• Both respond rather slowly to changes in light intensity. For
cadmium selenide, the response time (tres) is around 10
ms, while for cadmium sulfide it may be as long as 100 ms.
• Temperature sensitivity is another important difference
between the two materials.
• There is a large change in the resistance of a cadmium
selenide cell with changes in ambient temperature, but the
resistance of cadmium sulfide remains relatively stable.
 Solid-State Photo detectors
• Photodiodes
• Phototransistors
• PIN diodes
• Avalanche photodiodes
• Silicon photo multipliers
 Thermocouples
• Thermocouples are the most widely used detectors employed in
IR spectrophotometers.
• In these detectors, the signal originates from a potential
difference caused by heating a junction of unlike metals.
• They are made by welding together two wires of metals in such a
manner that they form two junctions.
• One junction between the two metals is heated by the IR beam
and the other junction is kept at constant temperature.
• Due to difference in work functions of the metals with
temperature, a small voltage develops across the thermocouple.
• The receiver element is generally blackened gold or platinum foil,
to which are welded the fine wires comprising the thermoelectric
junction.
• The other junction is shielded from the incident radiation.
Changes in temperature of the order of 10−6 °C can be detected.
 Bolometers
• Bolometers give an electrical signal as a result of the variation in
resistance of a conductor with temperature.
• It consists of a thin platinum strip in an evacuated glass vessel,
with a window transparent in the IR range.
• Irradiation by the IR beam produces an increase in resistance of
the metal strip, which is measured with a Wheatstone bridge.
• Usually, two identical elements are used in the opposite arms of
the bridge.
• One of the elements is placed in the path of the IR beam and the
other is used to compensate for the changes in the ambient
temperature.
• Alternatively, the platinum strips may be replaced with
thermistors, which show a negative thermal coefficient of
electrical resistance.
• Bolometer arrays have become the focus of most uncooled
detector development
Pneumatic detector

Figure: Golay pneumatic cell

 Construction and Working Principle
• The pneumatic detector described by Golay is used to measure the
intensity of IR radiation by following the expansion of a gas upon
heating.
• The Golay cell comprises a chamber containing xenon, a gas of low
thermal conductivity.
• It is sealed at its front end by a blackened receiver.
• The rear wall is a flexible membrane with a mirrored surface on its rear
side.
• IR energy falling on the receiver warms up the gas in the chamber.
• A rise in temperature of the gas in the chamber produces a
corresponding rise in pressure and therefore, a distortion of the mirror
diaphragm.
• Light from a lamp inside the detector housing can be focused on the
diaphragm, which reflects the light on to a photocell.
• Movements of the diaphragm corresponding to the amount of incident
IR energy changes the incident light energy on the photocell surface
and causes a change in the photocell output.
• By periodically interrupting the incident radiation with a chopper, an AC
signal is produced by the photocell which can be amplified.
• Another form of pneumatic detector is capacitor
microphone type in which the varying expansion
of the gas affects the capacitor film, which in turn
produces the variation in the electrostatic
capacity.
• Pneumatic detectors function at all wavelengths
throughout the IR region (as far as 400 m using
a diamond window).
• They have a low response time.
• The whole receiver is sensitive to the radiations,
thereby eliminating the need for optical
alignment.
 Pyroelectric detectors
• Pyroelectric detectors certain types of crystals (ferroelectric) get
polarised in a well defined direction, known as the polar axis.
• Because the degree of polarisation is temperature dependent,
heating or cooling a slice of such a crystal will create an
accumulation of charge (on the faces normal to the polar axis),
that is, proportional to the variation in polarisation caused by the
temperature change.
• This is the pyroelectric effect.
• Temperature fluctuations produce a charge change on the surface
of pyroelectric crystals, which produces a corresponding electrical
signal. This temperature gradient can be created by the
absorption of light.
• A pair of electrodes normal to the polar axis of the crystal may be
used to measure the voltage generated within the crystal due to
temperature changes.
• Pyroelectric detectors are mostly made from single crystal
triglycine sulphate (TGS).
Figure: Pyroelectric detector
• A pyroelectric detector contains a piece of ferroelectric crystal
material with electrodes on two sides – essentially a capacitor.
• One of those electrodes has a black coating (or a processed absorbing
metal surface), which is exposed to the incident radiation.
• The incident light is absorbed on the coating and thus also causes
some heating of the crystal, because the heat is conducted through
the electrode into the crystal.
• As a result, the crystal produces some pyroelectric voltage.
• The voltage generated is usually applied to a field effect transistor,
which is an integral part of the detector package.
• Pyroelectric detectors are current sources with an output that is
proportional to the rate of change of their temperatures.
• Pyroelectric materials absorb quite strongly in the FIR and have
essentially flat wavelength response from the near IR through the
FIR.
• Though the pyroelectric detectors have low sensitivity as compared to
Golay detector, they are preferred in the FIR range due to faster
response.
 Types of IR Spectrophotometers

• Optical Null Method

• Ratio recording
• The IR radiation is passed simultaneously through two
separate channels, one containing the sample, the
other the reference.
• The two beams are re-combined into a common axis
and are alternately focused on the detector.
• If the intensities of the sample and reference beams
are exactly equal, then no alternating intensity
radiation goes through the slit.
• If the sample absorbs some radiation, an alternating
intensity radiation is observed by the detector, which
produces an AC signal.
• This AC signal can be selectively amplified in a tuned
amplifier.
• As the amplifier is tuned to the chopping frequency of
one light beam, signals of frequency different from
the chopping frequency are not amplified.
• The alternating signal from the detector is used to drive a
servomotor, which is mechanically coupled to an optical
wedge or a fine-toothed comb attenuator.
• Movement of the comb occurs when a difference in power
of the two beams, is sensed by the detector.
• The motor will stop when the reference and the sample
beam intensities are exactly equal.
• The movement of the motor is synchronised with the
recorder pen, so that its position gives a measure of the
relative power of the two beams and thus the transmittance
of the sample.
• The record obtained, therefore, is of the sample absorption
as a function of spectral frequency.
• The teeth of the attenuator comb are accurately cut, so that
a linear relationship exists between the lateral movement of
the comb and the decrease in power of the beam.
Recording-type double-beam optical-null principle
IR spectrophotometer

Figure: Block diagram of a double-beam IR spectrophotometer

 Working Principle
• The two light beams from light source pass through the sample and
reference cells.
• With a switching frequency of 12.5 Hz, the rotary sector mirror
alternately conducts the radiation of the sample and reference
channels into the monochromator.
• Here, the light of the respective wave number is reflected and passed
on to the thermocouple, which delivers a 12.5 Hz AC voltage signal,
when the intensities of the two optical paths are not equal.
• After amplification in the pre-amplifier and main amplifier and after
phase-sensitive rectification in the demodulator, this AC voltage signal
controls the servomotor, which adjusts the attenuator diaphragm in
the reference channel until the intensities of both beams are equal
and the signal disappears.
• The servomotor also drives the recording pen, which records the
sample’s transmittance corresponding to the position of the
attenuator diaphragm.
 Ratio Recording Method
• In the direct ratio recording system, radiation from the
source is passed alternately through two separate channels,
one passing through the sample cell and the other through
the reference cell.
• The two signals from the detector are amplified, rectified
and recorded separately.
• The sample signal is displayed as a proportion of the
reference signal.
• In this method, attempts are not made to have a physical
equalisation of the sample and reference beam intensities.
• The reference signal is used to drive the slit width control,
so that the energy level reaching the detector is constant.
• This system is, however, not favoured in IR instruments, as
it requires a very stable amplifier system and the chopping
system.
Fourier Transform Infrared
Spectroscopy
 Fourier Transform Infrared Spectroscopy-
Introduction

• The Fourier transform technique depends upon the

basic principle, that any wave function could be
represented as a series of sine and cosine
functions with different frequencies.
• The determination of the sine and cosine
components of a given wave function is known as
Fourier transformation (FT).
 FTIR spectrometers
• In order to obtain spectra of the samples, a spectrometer
must be able to separate the optical frequencies of the IR
light after it passes through the sample.
• Dispersive spectrometers separate the optical frequencies
spatially using a prism or a diffraction grating before the
light passes through the sample.
• FTIR spectrometers modulate the IR beam before passing
through the sample.
• This is done by the interferometer which causes each IR
frequency to be modulated with a unique frequency of
modulation.
• After the IR beam passes through the sample, the radiation
intensity is detected and the frequencies are demodulated
via a FT.
• The heart of a FTIR spectrometer is a two-
beam interferometer, most commonly of the
Michelson type.

Figure: Principle of Michelson interferometer

 Working of Interferometer
• The radiation entering the interferometer is split into two beams
by a beam splitter.
• Beam A follows the straight path before returning to the beam
splitter, whereas the distance travelled by beam B can be varied,
before it re-combines with beam A.
• When the beams A and B recombine, an interference pattern is
produced, which is incident on a detector. When the two beams
are in phase at the beam splitter, maximum intensity will reach
the detector.
• The intensity will be minimum in case the beams are out of phase.
• If provision is made in such a way that mirror M2 is displaced
uniformly, the detector output will be a sine wave, whose
frequency is determined by the translation velocity of M2 and the
wavelength of the monochromatic radiation, and the amplitude of
the signal will depend upon the intensity of incoming radiation.
• If the incoming radiation is polychromatic, the detector output wave of
unique frequency is produced for each component.
• The overall detector output as a function of time, will be the sum of the
waves for each frequency component.
• The resulting signal is called an interferogram which has the unique
property that every data point (a function of the moving mirror position)
which makes up the signal has information about every IR frequency
which comes from the source.
• This means that as the interferogram is measured, all frequencies are
being measured simultaneously.
• Because the analyst requires a frequency spectrum (a plot of the intensity
at each individual frequency) in order to make an identification, the
measured interferogram signal cannot be interpreted directly.
• A means of ‘decoding’ the individual frequencies is required. This can be
accomplished via a well-known mathematical technique called the Fourier
transformation.
• This transformation is performed by the computer which then presents the
user with the desired spectral information for analysis.
Major Components of FTIR
Spectrophotometer

Figure: Principle of FTIR Spectrometer

• The Source: IR energy is emitted from a glowing black body
source. This beam passes through an aperture which controls the
amount of energy presented to the sample and to the detector.
• The Interferometer: The beam enters the interferometer where
the “spectral encoding” takes place. The resulting interferogram
signal then exits the interferometer.
• The Sample Compartment: The beam enters the sample
compartment where it is transmitted through or reflected off of
the surface of the sample, depending on the type of analysis
being accomplished. This is where specific frequencies of energy,
which are uniquely characteristic of the sample, are absorbed.
• The Detector: The beam finally passes to the detector for final
measurement. The detectors used are specially designed to
measure the special interferogram signal.
• The Computer: The measured signal is digitised and sent to the
computer where FT takes place. The IR spectrum is then displayed
user for interpretation and any further manipulation.
Figure: Block diagram of a typical FT spectrometer
• The source and detector are the conventional ones that
are used in IR spectrometry.
• A small computer is employed to control the scan
system and carry out the mathematical transformation.
• The heart of the system is the mirror scan mechanism.
• For faithful reproduction of the spectrum from the
interferogram, the detector output must be known as a
function of mirror displacement.
• The measurement of mirror displacement becomes
increasingly difficult, as the wavelength decreases.
• At shorter wavelengths, a special technique known as
fringe referencing technique is used.
• Here, one transducer is used to drive simultaneously the
movable mirrors of two interferometers through identical
displacements.
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Figure: Constructive and destructive interferogram

• Sample radiation passes through one interferometer and
produces the sample interferogram.
• Reference interferogram is recorded using a He-Ne laser.
• This reference laser ensures that sampling of the detector
signal occurs reproduceably from scan to scan.
• Since the interferogram of a monochromatic source (laser)
is a sine wave, its zero crossings provide an accurate
trigger for digitised data collection of the main
interferogram.
• Further, use of He-Ne laser reference interferogram
provides an internal frequency calibration, important to
high resolution studies.
• The application of FT spectrometry has been extended to
uv/visible region in the entire spectral range from 160 nm–
700 nm.
• The resolution achieved is much greater than the
instruments based on grating monochromators.
 Advantages of FTIR
Some of the major advantages of FTIR over the dispersive technique
include:
• Speed: Because all of the frequencies are measured
simultaneously, most measurements by FTIR are made in a
matter of seconds rather than several minutes.
• Sensitivity: Sensitivity is dramatically improved with FTIR for
many reasons. The detectors employed are much more sensitive,
the optical throughput is much higher which results in much lower
noise levels, and the fast scans enable the co-addition of several
scans in order to reduce the random measurement noise to any
desired level.
• Mechanical Simplicity: The moving mirror in the interferometer
is the only continuously moving part in the instrument. Thus,
there is very little possibility of mechanical breakdown.
• Internally Calibrated: These instruments employ a He-Ne laser
as an internal wavelength calibration standard. These instruments
are self-calibrating and never need to be calibrated by the user.
UV – VIS spectrophotometer
 Introduction
• Ultraviolet-visible (UV-Vis) spectrophotometry is a
technique used to measure light absorbance across the
ultraviolet and visible ranges of the electromagnetic
spectrum.
• When incident light strikes matter it can either be
absorbed, reflected, or transmitted.
• The absorbance of radiation in the UV-Vis range causes
atomic excitation, which refers to the transition of
molecules from a low-energy ground state to an excited
state.
• Before an atom can change excitation states, it must
absorb sufficient levels of radiation for electrons to
move into higher molecular orbits.
• A UV-Vis spectrophotometer can use this principle to
quantify the analytes in a sample based on their
absorption characteristics.
• A UV-Vis spectrophotometer measures the intensity of
light transmitted through a sample compared to a
reference measurement of the incident light source.
• According to Beer-Lambert Law, the amount of light
absorbed is directly proportional to the concentration
of the sample and the distance the light travels
through the sample; the path length.
• UV-Vis spectrophotometers are able to determine the
concentration of specific analytes in a microvolume by
controlling the analysis wavelengths and the path
length.
• Since the UV-Vis range spans the range of human
visual acuity of approximately 400–750 nm, UV-Vis
spectroscopy is useful to characterise the absorption,
transmission, and reflectivity of a variety of
technologically important materials, such as pigments,
coatings, windows, and filters.

Figure: Single beam UV-VIS spectrophotometer

• The light source is usually a Tungsten-Halogen Lamp
or deuterium-discharge lamp for UV measurements
and a tungsten halogen lamp for visible and NIR
measurements.
• The instruments automatically swap lamps when
scanning between the UV and visible regions.
• The wavelengths of these continuous light sources are
typically dispersed by a holographic grating in a single
or double monochromator or spectrograph.
• The spectral bandpass is then determined by the
monochromator slit width or by the array-element
width in array-detector spectrometers.
• The detector in single-detector instruments is a
photodiode, phototube or PMT.
• UV-Vis-NIR spectrometers utilise a combination of a PMT
and a Peltier-cooled PbS IR detector.
• The light beam is redirected automatically to the
appropriate detector when scanning between the visible
and NIR regions.
• The diffraction grating and instrument parameters such as
slit width can also be change.
• The UV-Vis spectral range is approximately 190–900 nm, as
defined by the working range of typical commercial UV-Vis
spectrophotometers.
• The short-wavelength limit for simple UV-Vis spectrometers
is the absorption of ultraviolet wavelengths less than 180
nm by atmospheric gases.
• High-end commercial UV-Vis spectrophotometers extend
the measurable spectral range into the NIR region as far as
3,300 nm.
 Types of UV-Vis spectrophotometer

• Single beam and

• Double beam
Single beam spectrophotometer
 Working of Single beam spectrophotometer
• A spectrophotometer measures the light that passes through
the sample, to then strike the detector, where it is measured.
• Let’s call the amount of light at wavelength λ incident on the
sample cuvette Io and the amount of light exiting the sample
to hit the detector I.
• If we now calculate the percent of the ratio these two values
(I/Io)*100 we get a percent transmission (%T) value.
• If the sample transmits all the light at a given wavelength then
%T = 100; however, if the sample absorbs light then we will
have the case where I < Io and %T will be a number less than
100.
• If the beam is totally blocked or absorbed by the sample, then
%T = 0.
• When a spectrophotometer is turned on it is literally
as “dumb as a stump”. In other words, the instrument
has not been calibrated for the values of 100 %T or 0
%T.
• This 100 %T calibration is acquired by a process
called background correction for a spectrum. it is also
sometimes called auto-zero for a single wavelength
measurement.
• A background correction is performed by removing
the sample from the instrument and measuring the
amount of light that strikes the detector in the scan
range of all wavelengths in the spectrum.
• These 100 %T values (called a background scan) are
then stored in memory for use in calculating accurate
sample %T values for a sample.
Double beam spectrophotometer
• These instruments are configured to allow the automatic and
simultaneous (or near simultaneous) measurement of the sample
and reference beam intensities.
• In a double-beam system, the monochromatic light from the
monochromator is split into a sample beam (S) and reference
beam (R) before entering the sample compartment and hitting the
detector.
• This can be accomplished using a beam splitter to divide the beam
into the reference beam and sample beam.
• In the double-beam-in-space configuration, the intensities of the
split beams are measured simultaneously after passing through
the sample and reference cells.
• Alternately, in a double-beam-in-time spectrophotometer, the
beams pass through a modulator which allows the detector to see
either the reference beam or the sample beam.
• Since the source beam is directed through a reference cell part of
the time and through the sample cell the rest of the time, the
term ‘double-beam-in-time’ is used.
• The main disadvantage of single-beam
spectrophotometer is that the instrument settings
have to be adjusted to give a 100% reading with
the reference in the beam, before the sample is
examined.
• This drawback is overcome by using a double-
beam instrument, wherein the arrangement is
such that the radiation beam is shifted
automatically to pass alternately through the
sample and reference cuvettes.
• The cuvettes themselves are not shifted and
remain in their fixed position.
Thank you