Important Highlights

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2. WHAT DOES THE WORD MEAN
SPECTROFLOURIMETRY
 A device for measuring and recording
fluorescence spectra
 It is made of two words
SPECTRO + FLOURIMETRY
SPECTRO = ELECTROMAGNETIC SPECTRUM
FLOURIMETRY = EMISSION OF VISIBLE
RADIATION
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3. FLUORESCENCE
 When a beam of Light is incident on certain
substances they emit Visible Elecromagnetic Radiation
/ Light.
This Phenomenon is Fluorescence.
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4. LUMINESCENCE
 No. of Substances are also known which loses their
only part of their excess energy as Heat and Emit the
remaining energy as electromagnetic radiation of
wavelength longer than that absorbed
This process of Emitting Radiation is known as
Luminescence.
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5. PRINCIPLE
1. SINGLET AND TRIPLET STATES :
Simply it means –
“No. of Unpaired Electrons in the Absence of Magnetic
Field”.
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6. 2. Excited State ( Molecules / Atoms ) :
Three Possibilities Probably Can occur under following
conditions.
i. Excited Single State (Unstable).
ii. Molecule Emitting Visible Radiations
[Fluorescence].
iii. Molecule in Metastable Triplet State
[Phosphorescence].
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10. RADIATION SOURCE :
 MERCURY/ XENON ARC LAMP :
 Wavelength Range : 200 – 1800 nm
 Mostly Radiation of UV – Visible Spectra.
 High Intensity.
 Can be used to Very Dim Fluorescence.
 Excitation of Fluorescent Substances is Maximum.
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11. MONOCHROMATOR :
 Diffraction Grating Monochromators :
 It selects Very Selective Wavelength of light .
 Which Produce Excitation in the Molecules / Atoms.
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12. DETECTOR
 Photomultiplier Tubes :
 Consists of a photosensitive cathode, several dynodes
and a collection anode.
 Dynodes are responsible for the increase in signal by
electron multiplication.
 Fast response , Sensitive for detection of light .
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13. APPLICATIONS:
 1. Determination of Uranium in Salts by Fluorimetry.
 2. Inorganic ions do not exhibit Fluorescent. Exhibits
Fluroscence with Fluorescent Chelates.
 3. Determination of Aluminium [ iii ] Alloy form
complex with Pantachrome Blue- Black Dye.
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14.  4. Estimation of Calcium by Flourimetry with Calcein
Solution .
 5. Determination of Vitamin B1 [ Thiamine].
 6. Determination of Vitamin B2 [ Riboflavin].
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