UV-VISIBLE SPECTROSCOPY

UV-visible spectroscopy refers to absorption spectroscopy in the ultraviolet-visible

spectral region.
PRINCIPLE
Beers Law:
For monochromatic radiation, absorbance is directly proportional to the path length b
through the medium and the concentration c of the absorbing species.
A bc
A= bc
Where,
c is the concentration in moles per litre
b is the cell length in centimeters
is the molar absorptivity in L/mol/cm.
INSTRUMENTATION
Instruments used for measuring the absorption of ultraviolet and visible radiation are made
up of the following components
1. Source
2. Wavelength selectors
3. Sample containers
4. Radiation transducers
5. Signal
processors and readout devices
SOURCE

WAVELENGTH
SELECTOR

SAMPLE

DETECTOR

READOUT
DEVICE

SOURCE
For molecular absorption measurements, a continuum source is required, whose radiant
power does not change sharply over a considerable range of wavelengths.
DEUTERIUM AND HYDROGEN LAMPS
A continuum spectrum in the ultraviolet region is produced by the electrical excitation of
deuterium or hydrogen at low pressure.
The mechanism by which a continuum spectrum is produced involves the initial
formation of an excited molecular species, which dissociates to give two atomic species
and an ultraviolet photon.
The consequence is a true continuum spectrum from about 160 nm to the beginning of
the visible region.
In low voltage deuterium lamps (operating at 40V), an arc is formed between a heated
oxide filament and a metal electrode.
This heated filament provides electrons to maintain a direct current. A regulated power
supply is required to obtain a constant light intensity.
Both hydrogen and deuterium lamps produce outputs in the range of 160-800 nm.

 In the UV region (190-400 nm) a continuum spectrum exists, but at longer wavelengths
(>400nm), the spectrums no longer a continuum.
Quartz windows must be employed in deuterium and hydrogen lamps as glass absorbs
strongly at wavelengths less than 350nm.
The useful lower limit of deuterium lamps is 190nm.

TUNGSTEN FILAMENT LAMPS AND TUNGSTEN HALOGEN LAMPS

Tungsten filament lamps are the most common source of visible radiation.
The operating filament temperature is 2870K.
These lamps are useful for the wavelength region between 350 and 2500 nm.
Tungsten halogen lamps, also called as quartz halogen lamps, contain a small quantity of
iodine in a quartz envelope. This envelope also houses the tungsten filament.
Quartz allows the filament to be operated at a temperature of about 3500K.
As a result, tungsten (W) formed by sublimation reacts with iodine to form a volatile
compound, WI2
When the molecules of this compound strike the filament, the compound decomposes and
tungsten is re-deposited.
Hence these lamps are highly efficient and extend their range into the UV region as well.

XENON ARC LAMPS

These lamps produce intense radiation by passage of current through an atmosphere of
xenon.
A continuum spectrum is obtained, with the wavelength range of 200 to 1000nm.
Peak intensity occurs at 500nm.
2. WAVELENGTH SELECTORS
They are used to select a particular wavelength. For UV-visible spectroscopy, filters or
monochromators can be used. In gratings, echellete gratings are specifically used.
FILTERS
There are two types of filters - interference filters and absorption filters.
INTERFERENCE FILTERS
They rely on optical interference to provide narrow bands of radiation.
They consist of a transparent dielectric layer, present between two semi transparent
metallic films, sandwiched between two plates of glass.
When a perpendicular beam of radiation strikes this array, fraction passes through the
first metallic plate and the remainder is reflected.
A similar partition takes place when it strikes the second metallic film.
If the reflected portion is of proper wavelength, it is partially reflected from the inner side
of the first layer and this particular wavelength is reinforced.
ABSORPTION FILTERS

 They are used for band selection in the visible region and are less expensive than
interference filters.
They function by absorbing selected portions of the spectrum.
These filters consist of a coloured glass, or a dye suspended in gelatin, sandwiched
between glass plates.
They have effective bandwidths that range from 30 to 250 nm.
MONOCHROMATORS
They are used to continuously vary the wavelength of radiation over a broad range.
The components of a monochromator are - entrance slit, collimating lens, a prism or
grating, a focusing element and an exit slit.
Two types of dispersing elements are found in monochromators - reflection gratings and
prisms.
PRISM MONOCHROMATORS
Prisms can be used to disperse UV, visible and infrared radiation.
There are two types of prism - 30prism and 60prism.

60 prisms made from a single block of material. If crystalline quartz is used for
construction, two 30 prisms are made and cemented together.
Hence there is no net polarization and this type of prism is called Cornu prism.
The Littrow prism is a 30 prism that has a mirrored back.
Here refraction takes place twice at the same interface.
GRATING MONOCHROMATORS
Gratings consist of a hard, optically flat, polished surface, with a large number of parallel,
closely spaced grooves.
There are two major types of gratings - reflection gratings and transmission gratings.
ECHELETTE GRATING
This grating consists of a grooved or blazed surface.
It has broad faces from which reflection occurs and narrow unused faces.
This geometry provides a highly efficient diffraction of radiation.
The reason for blazing is to concentrate radiation in a particular direction.

- The other types of gratings are concave gratings and holographic gratings.
3. SAMPLE CONTAINERS
The cells or cuvettes must be constructed of a material that passes the radiation, in the
spectral region of interest.
For UV region (below 350 nm), quartz or fused silica is required.
Silicate glasses can be employed in the region between 350 and 2000 nm.

Plastic containers are also used in the visible region.

The most common path length for studies in UV and visible region is 1 cm.
Cylindrical cells are employed because they are inexpensive.
The quality of absorbance data depends upon the way the cells are used and maintained.
Fingerprints, grease and other deposits alter the transmission characteristics of the cell.
Hence the cells should be thoroughly cleaned before and after use and must be regularly
calibrated.

4. RADIATION TRANSDUCERS
Photon transducers are used in UV-visible spectroscopy.
BARRIER LAYER-PHOTOVOLTAIC CELLS
Photovoltaic cells are simple devices used to detect and measure radiation in the vivible
region.
These cells have maximum sensitivity at 550 nm.
It consists of a flat copper or iron electrode on which a semiconductor layer (E.g.
selenium) is deposited.
The outer layer of the semiconductor is coated with a thin transparent metallic film of
gold or silver, which serves as the second electrode. The entire array is protected by a
transparent envelope.
When radiation of sufficient energy reaches the semiconductor, electrons and holes are
formed.
The electrons move towards the metallic film and then migrate through the external
circuit, to interact with the holes that move towards the semiconductor base.
This results in an electrical current of magnitude, proportional to the number of photons
that strike the semiconductor surface.
Currents of the order 10 to 100 A are typical.
PHOTOMULTIPLIER TUBES
They are sensitive to ultraviolet and visible radiation and used for the measurement of
low radiant powers.
The photocathode surface is coated with a photo emissive material and it emits electrons
when exposed to radiation.
Nine additional electrodes called dynodes are present.
The dynode D1 is 40V more positive than the cathode; hence electrons are accelerated
towards it.
Each photoelectron causes the emission of several other photoelectrons.
These electrons are in turn accelerated towards the dynode D2, which is 90V more
positive than the dynode D1.
10^6 to 10^7 electrons are formed for each incident photon.
This cascade of electrons is finally collected at the anode; the resulting current is
converted to voltage and then measured.
5. SIGNAL PROCESSORS AND READ OUT DEVICES

Alphanumeric devices directly give the output of the measurements. The output is
displayed on an LCD or an LED screen.
TYPES OF UV-VISIBLE SPECTROSCOPIC INSTRUMENTS
There are three types of UV-visible spectroscopic instruments.
1. Single beam instrument
2. Double beam in space instrument
3. Double beam in time instrument
FOR ABSORPTION MEASUREMENTS,
SOURCE
A tungsten or deuterium lamp
WAVELENGTH SELECTOR
A filter or a monochromator for wavelength selection
SAMPLE CONTAINERS
Cuvettes that can be placed along the radiation beam
TRANSDUCERS
Photon or thermal
AMPLIFIER
READOUT DEVICES
computer
SINGLE BEAM UV-VIS SPECTROPHOTOMETER
Source
Battery-operated tungsten bulb with stabilized voltage supply
Wavelength Selector
Glass filters
Sample Containers
Test tubes
Transducers
Photovoltaic cell
Amplifier
Readout Devices
Analog meter
Merits
Simple and least expensive
DOUBLE BEAM UV-VIS SPECTROPHOTOMETER
Sophisticated, computer-controlled instruments with a range of 200 to 1000 nm or more.
Source
Wavelength Selector
Sample Containers
Transducers
Amplifier
Readout Devices
Merits

Interchangeable Tungsten And Deuterium Lamp Sources,

High-Resolution Grating Monochromator With Variable Slits
Rectangular Silica Cells
Photomultiplier Tubes
Digitized, Processed Computer
Highly Sophisticated Producing More Accurate Results

DOUBLE BEAM IN-SPACE

Here two beams are formed in space by a V-shape mirror called a beamsplitter.
One beam passes through the reference solution to a photodetector.
Second beam simultaneously pass through the sample to a second detector.
The two outputs are amplified, and their ratio is determined electronically or by a computer and
displayed by the readout device.
The measurement is a two-step operation involving first the zero adjustment with a shutter in
place between selector and beamsplitter. Then the shutter is opened and the transmittance or
absorbance is displayed directly.

DOUBLE BEAM IN-TIME

ROTATING SECTOR MIRROR

It is motor-driven and is made up of pie-shape segments

Half of which are mirrored and half of which are transparent.
The mirrored sections are held in place by blackened metal frames that periodically interrupt the
beam and prevent its reaching the transducer. This interruption period is used to perform the
dark-current adjustment.
WORKING
Here, the beams are separated in time by a rotating sector mirror that directs the entire beam
from the monochromator first through the reference cell and then through the sample cell.
The pulses of radiation are recombined by grid mirror, which transmits one pulse and reflects the other
to the transducer. Now the ratio of two radiations are calculated, amplified and then shown in read out
device.