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Fluorimetric Analysis. Spectroscopy. Fluorimetric Analysis : - PowerPoint PPT Presentation
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Fluorimetric Analysis
Absorption of energy
Emission of energy
Spectroscopy
Absorption of energy
Electronic transition UV/Visible
VibrationIR
Rotationmicrowave
Emission of energy
Flame emission Photoluminescence
Photoluminescence
FluorescenceThe excited molecule
returns to the ground state immediately after excitation
and vibration relaxation
PhosphorescenceThere is a time delay
between the excitation and returning to the ground
state after vibration relaxation
Fluorimetric Analysis : method of chemical analysis in which a sample,
exposed to radiation of one wavelength, absorbs this radiation and reemits radiation of the same or longer wavelength ,the intensity of reemited radiation is almost directly proportional to the concentration of the fluorescing material. Also known as fluorescence analysis; fluorometry.
Flurometer SpectrophotmeterSource of light Xenon arch lamp
Or Hg vapor lamp-Perpendicular to sample
-Tangeston.-Deturium- Stright line.
Detector -Two monochromator-Very sensitive
- One monochromator
Sample Cell -Cuvette completely transberent - cell compartment is painted blank
- Only two side transperent
Comparison
Objectives:1- Determination of excitation and emission spectra of
Quinine SO4.
2- Prepare of calibration curve of Quinine SO4.
3-Determination of Concentration of unk. Quinine SO4 solution.
4-Demonstrate Quenching effect of KI on flourescence of quinine SO4.
Quenching effect
What does it mean by Quenching ?Quenching refers to any process which
decreases the fluorescence intensity of a given substance.
Quenching may occur by several mechanism:
collisional or dynamic quenching static quenching quencing by energy transfer charge transfer reactions
Quencher
• Chemical Quencher as alkali halide
• Self Quenching ??