UV Spectrophotometer

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Instrumentatio n
Components of spectrophotometer
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Source Monochromator Sample compartment Detector Recorder

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SAMPLE

RADIANT SOURCE

WAVELENGTH SELECTOR

SOLVENT

DETECTOR

PHOTO -

READOUT

Fig.-block diagram of instrumentation of UVspectrophotometer

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Entrance slit

Exit slit

Light source a)D2 Lamp b)WI Lamp

monochromat or

sample

detecto r

amplifier

Read out

Fig.- block diagrammatic representation of UVspectrophotometer

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Conventional Spectrophotometer

Schematic of a conventional single-beam spectrophotometer

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Conventional Spectrophotometer

Optical system of a double-beam spectrophotometer

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Light source

Distribution of energy through spectrum is function of temperature. For Visible regionTungsten filament lamp Use for region 350nm to 2000nm. Problem-

Due to evaporation of tungsten life period decreases. It is overcome by using tungsten-halogen lamp. Halogen gas prevents evaporation of tungsten.
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q q

For ultra violet regionHydrogen discharge lamp consist of two electrode contain in deuterium filled silica envelop. gives continuous spectrum in region 185-380nm. above 380nm emission is not continuous. UV-Vis spectrophotometer have both deuterium & tungsten lamps. Selection of lamp is made by moving lamp mounting or mirror to cause the light fall on monochromator.
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Deuterium lamps:Radiation emitted is 3-5 times more than the hydrogen discharge lamps. Xenon discharge lamp:Xenon stored under pressure in 10-30 atmosphere. It possesses two tungsten electrode separated by 8 cm. Intensity of UV radiation more than hydrogen lamp. Mercury arc:Mercury vapour filled under the pressure .
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Monochromat or
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Filters

a)Glass filters

Made from pieces of colored glass which transmit limited wavelength range of spectrum. produced by incorporation of oxide of vanadium, chromium, iron, nickel, copper. band width 150nm.
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Color

Wide

PrismsPrism bends the monochromatic light. Amount of deviation depends on wavelength. Quartz prism used in UV-region. Glass prism used in visible region spectrum. They produce non linear dispersion.

Function

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Fig.-Mechanism of prism working

Fig.-mechanism of working of prism.

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GratingLarge number of equispaced lines on a glass blank coated with aluminum film.
q

Normal surface vector

Normal to groove

Blaze angle
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Spectroscopy requires all materials in the beam path other than the analyte should be as transparent to the radiation as possible. qThe geometries of all components in the system should be such as to maximize the signal and minimize the scattered light. qThe material from which a sample cuvette is fabricated controls the optical window that can be used. Some typical materials are: Optical Glass - 335 - 2500 nm Special Optical Glass 320 - 2500 nm Quartz (Infrared) 220 - 3800 nm Quartz (Far-UV) 170 - 2700 nm Keep the cuvette clean. Dont clean with paper products. Store dry. Dont get finger prints on them. Store carefully and gently 4/14/12
q

Sample cell (cuvette)

Detectors
q

Three common types of detectors are used Barrier layer cells Photocell detector Photomultiplier Photo voltaic cells or barrier layer cells :They are primarily used for measurement of radiation in visible region. Maximum sensitivity-550nm. It consist of flat Cu or Fe electrode on which semiconductor such as selenium is deposited. on the selenium a thin layer of silver or gold is
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A barrier exist between the selenium & iron which prevents the electron flowing through iron. Therefore electrons are accumulated on the silver surface. These electrons are produced voltage. terminal Silver surface seleni um + termina fig.-Barrier l layer cell 4/14/12

Photocell detector:It consist of high sensitive cathode in the form of a half cylinder of metal which is evacuated. Anode also present which fixed along the axis of the tube Photocell is more sensitive than photovoltaic cell. lig ht

Fig.- photocell detector

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Photomultiplier tube:it is generally used as detector in UVspectrophotometer It is the combination of photodiode & electron multiplier. It consist of evacuated tube contains photo-cathode. 9-16 electrodes known as dynodes.

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Recorder

Signal from detector received by the recording system The recording done by recorder pen.

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q q

Description of UVspectrophotometer

Single beam spectrophotometer:Double beam spectrophotometer:-\ Advantage of double beam spectrophotometer:It is not necessary to continually replace the blank with the sample or to adjust the autozero. The ratio of the powers of the sample & reference is constantly obtained. It has rapid scanning over the wide wavelength region because of the above two factors.
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