Instrumentation of

Absorption
Spectroscopy
The adsorption of radiation in a solution is
quantitatively measured by using
Photoelectric colorimeters (Vis)
 Spectrophotometers (Uv, Vis, IR)

Principle: When a narrow band of wavelengths is

incident on the absorbing sample, part of light is
absorbed and rest is transmitted. The amount of light
absorbed depends on the concentration and path
length travelled by light through the solution. Thus it
is governed by Lambert-Beer’s law, A=ϵcl
 Photometer Spectrophotometer
1. Works in visible range 1. Works in UV, visible and
IR range
2. Use color filters
2. Use monochromatic
3. Allows band of radiations
wavelength of
3. More accurate
complimentary colors
4. Band width is minimum
of sample to be incident
5. Lambert beer’s law is
4. Quantitaive analysis of
followed
coloured compounds
6. Structural determination
5. Cheaper as compared alongwith quantitative
to spectrophotometers analysis
 Types of Photoelectric colorimeters

• Single beam photoelectric

colorimeter ( one optical beam)

• Double beam photoelectric

colorimeter (two identical optical beam)
 Single beam photoelectric colorimeter

•Narrow band of wavelength is incident on the absorbing

sample.
•All measurements are done using a single optical path for the
beam of light

•The absorbance (% transmittance) are determined after

adjusting the absorbance of the solvent to 0 (100% T).
Double beam photoelectric colorimeter
Advantages of Single Beam
• Single beam instruments are less
expensive
• High energy throughput due to
non-splitting of source beam
results in high sensitivity of
detection
Disadvantages
• Instability due to lack of
compensation for disturbances
like electronic circuit fluctuations,
voltage fluctuations, mechanical
component’s instability or drift in
energy of light sources. Such
drifts result in abnormal
fluctuations in the results.
Advantages of Double Beam
• Modern improvements in
optics permit high level of
automation and offer the
same or even better level of
detection as compared to
earlier single beam systems.
• Instability factors due to
lamp drift, stray light, voltage
fluctuations do not affect the
measurement in real-time.
• Error due to solvent or
impurity is balanced out
• Readings are not affected by
change in sensitivity of
photocells as null method is
used
 Components of photoelectric colorimeter
1. Source of visible radiation: Tungsten filament is mostly used for
obtaining continuous radiations in visible region (400-750 nm)

2. Lens: to obtain parallel beam of light

3. Filter or wavelength selector: for required wavelength to pass

through

4. Detector/Transducer: to convert transmitted beam emerging

from sample into electric current. Its a type of photovoltaic cell

5. Recorder: It records the output of detector as absorbance or %T

in digital form.
 Working
• Filter which gives maximum absorbance with
sample solution is selected
• Cuvette is first filled with pure solvent and %T is
adjusted to 100% (or A to 0) (diaphragm or rheostat is
adjusted accordingly)
• Then cuvette is filled with sample solution
without disturbing the diaphragm or rheostat
• Absorbance of sample is noted and
concentration can be found using Lamberts
beer’s law
A= ϵ c l
 Spectrophotometer requires
• A Stable and cheap radiant energy source
• A monochromator, to break the polychromatic
radiation into component wavelength (or)
bands of wavelengths
• Sample cells (curettes), to hold the sample
• A Photosensitive detector to convert radiant
energy into electrical signal
• Associated readout system
• Spectrophotometers are refined photoelectric
colorimeters, there incident radiations is
nearly monochromatic
• In visible spectrophotometers, white light is
resolved into its components and suitable
wavelength is selected
• While spectrophotometers allow continuous
choice of wavelength
Entrance slit

Objective lens

Exit slit
• Optical system is separated from electronic
circuitry by a wall
• Wall act as a shield for stray light and permits
error-free operation
• For Uv spectrophotometers
optics: quartz
• Stable regulated power supply is required to
power the lamp
(6-V storage battery as voltage source)
 Source
Source should have readily detectable output
radiation for desired wavelength
For visible region:
 Tungsten filament incandescent lamp
useful in near UV (325 nm) to near IR (3µm)
For UV region:
 Hydrogen/ deuterium lamp
used from 185 nm to 375 nm
Source has UV transparent quartz windows and to
be cooled regularly to dissipate generated heat
 Monochromators
• It is used to get radiation of high spectral
purity of any desired wavelength from a
continuous source of radiation

They are of two types

1) Prism monochromator
2) Grating monochromator
 Prism monochromator

For visible region : prism & lens are made up of glass

For UV region : prism & lens made up of quartz

 Grating monochromators

• Grating disperse radiations uniformly throughout the

entire spectrum
• Advantage being dispersion along the focal plane of
the exit slit is independent of wavelength
• So for given setting of slit width, same band width is
obtained regardless of spectral region
 Transmission
Reflection Gratings
Gratings
• Made by ruling a • Made by ruling a
large no. of parallel large no. of parallel
lines on highly lines on glass
polished surface surface
such as Al

For UV & Vis

(15,000 to 30,000 lines/inch)
 Reflection grating

• Light which strikes the ruling is scattered and

dissipated while light which strikes the
unruled portion is reflected regularled.

• Relected waves produce an interference

pattern resulting in dispersion of light into its
components wavelength
 Transmission grating

• When light falls on transmission grating, each

groove act as individual source of light

• The transmitted rays from groove, produce an

interference pattern resulting in dispersion of
light into its components wavelength
 Advantages of grating
monochromators over prisms
• Grating gives better dispersion of light than
prisms

• Gratings are made up of non-corrosive material

• Not easily attacked by moisture

• Gratings can be used for a longer wavelength

range compared to prisms
 Optical Filters
• Glass filters: narrow band-pass filters, sharp cut filters
(colored by dispersing a pigment/dye in glass)

Narrow band-pass filters: dye absorbs all the

radiations expect the desired

Sharp cut filters: dyes absorbs all radiation Upto

specified wavelength, and pass radiation at longer
wavelength
 Optical Filters
• Interference filters: consists of two layers of
glass, inner surfaces have thin semi-transparent
films of metal such as silver and inner layer of
transparent material such as quartz or calcium
fluoride.
 Optical Filters

• Light undergoes interference at silver surface

• The unwanted wavelengths are removed by
selective reflection and required wavelength
can pass out of filter.
• Bandwidth of filter decreases as transmitted
radiation increases
• Interference filters gives narrow bands as
compared to glass filters
 Sample Cell
• Cell holding the solution must be transparent
• Cuvette with 1cm thick

UV region : Quartz
Visible region: quartz/glass
 Detector
• Function is to convert transmitted radiation
emerging from sample into an electric current

• Requirements of detector are:

1. It must respond to radiant energy over broad
wavelength range
2. Should be very sensitive
3. Must respond rapidly
4. Should give signal which is directly proportional
intensity of radiation
 Commonly used Detectors

1. Photoemissive cells

2. Photomultiplier tube

3. Photovoltaic/barrier cell
•Glass bulb is fitted with semicircular photocathode made up
of Cs/ metal coated with Cs/ mixture of K+ AgO
•Metal ring act as anode
 Current is amplified Output is
by electronic means conned to
calibrated
(taken as amount of meter in either
Radiation is light striking the T or A
incident on photocathode)
cathode

Flow of electron decrease

Photoelectrons
internal resistance which
are emitted
is proportional to current

Attracted to Current flows

anode in outer circuit
 Photomultiplier tube

•These are Photoemissive cells in which primary electrons (from

photocathode) falls on another Photoemissive cathode ejecting
secondary electrons from it
• Increasing positive potential are applied
•10 secondary electrons are emitted for every primary electron
 Photovoltaic/barrier cell
Transmitted light from sample

Emits electrons from Se surface

Electrons pass through hypothetical

barrier between Se and Ag collector
electrodes

Collected by silver electrode

• Under the influence of light, cell is formed
with iron plate as positive electrode and
metal ring ad negative electrode.
• This type of cell generates its own emf and
no external power supply is required
• When connected to galvanometer, current
will flow provided resistance is small
• Electric current produced is proportional to
amount of light falling on Se surface