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Name : J Srinivasa Rao
Designation : Lecturer
Branch : D.A.E.I.E
Institute : Govt. Polytechnic, Kothagudem
Year/Semester : VI Sem
Subject : Analytical Instrumentation
Subject Code : AEI-603
Topic : Analysers
Duration : 100 Min
Sub Topic : Spectrofluriometers
Teaching Aids : PPT, Pictures

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On completion of this period you will be able to

know about

• Spectrofluriometer

• Principle

• Construction and Working

• Applications

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• Spectroscopy

• UV / Visible spectrophotometer

• I.R. Spectrophotometer

• Flame Photometer

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• When a quanta of light impinges on a molecule, it is
absorbed and an electronics transition to a higher
electronic state takes place.

• The higher energy state is unstable and returns back to

the ground state by emitting light energy.

• If this transition takes 10-8sec then the light emitted is

called fluorescence light.

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Construction & Working of
• Spectrofluriometers uses two Monochromators, one to
supply selectively the excitation radiation and the other
to isolate and analyze fluorescence emission.

• The source of light is a 150W high pressure xenon arc


• The ellipsoidal mirror collects radiation from this lamp

and focuses on the entrance slit of the excitation
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• Monochromatic radiation of specific wavelength is
selected by angular positioning of the grating.

• A desired band pass is achieved by the selection of

appropriate interchangeable slits for the entrance and
exit positions of the monochromator.

• The radiant energy emerging from the exit slit of the

excitation monochromator activates specific molecules
within the sample resulting in fluorescent emission.

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• The emitted radiation is analyzed by the analyzing
monochromator, which permits only characteristic
sample emission to reach the detector, blocking all
undesired spectral regions.

• When fluorescent light falls on the photomultiplier, the

registered photocurrent can be indicated on the micro
ammeter, recorder or oscilloscope

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• In clinical laboratories for studying of adrenalin and non

adrenalin in urine and blood in screening tests for tumors
of adrenal glands.

• For testing of lactic dehydrogenates, steroids, DNA,


• For analysis of air pollution

• Used in agriculture and food industries to determine

insecticide and residue studies.

• To study the diffusion of air and water by sanitation

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• Spectrofluriometers is based on the principle of


• The light source used is Xenon arc lamp

• The monochromator used is Grating

• The detector used is photomultiplier tube

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1) The Principle of Spectroflurometer is based on
the principle of
a) Phosphorescence
b) Fluorescence
c) Chemiluminescence
d) All of the above

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2) The fluorescence light is emitted when the
energy transition from a higher energy state to
lower energy state takes place in a time of
 10-6
 10-8
 10-12
 10-14

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3) The detector used for Spectrofluriometers is
1. Photomultiplier tube
2. Bolometer
3. Thermal conductivity detector
4. All the above

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4) The source used for Spectrofluriometers is
a) Deuterium lamp
b) Tungsten lamp
c) Xenon arc lamp
d) Mercury lamp

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Frequently asked questions

1) Explain the principle construction and working of


2) List out the applications of Spectrofluriometers.

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