Spectroflurometry.pptx

Zoology

1. Spectrofluorometry Dr. G. D. HANDE Assistant Professor Shri. Shivaji Science College Amravati.

2. A spectrofluorometer is an instrument which takes advantage of fluorescent properties of some compounds in order to provide information regarding their concentration and chemical environment in a sample. A certain excitation wavelength is selected, and the emission is observed either at a single wavelength, or a scan is performed to record the intensity versus wavelength, also called an emission spectra

3. Principle: Spectrofluorometry, as the name suggests takes the advantage of the fluorescent properties.  So, before understanding about spectrofluorometry, it is necessary to know what is fluorescence. When a molecule after absorbing radiations, emits radiation of a longer wavelength, then this phenomenon is referred to as “ fluorescence .” Because of this, the compound absorbing in ultraviolet range might emit radiation in visible range. 

4. After understanding the basic principle of fluorescence, we will now come to the main principle of spectrofluorometry. As we have learnt in Chemistry, when an atom or molecule absorbs radiation, the energy of the photon absorbed lifts an electron to a higher orbital. Now, the electron needs to come down back to its ground state.

5. Instruments for Measuring Absorption of Light….

6. Fluorescence and Phosphorescence Excitation Beam Emitted Beam Detector

7. Filter Fluorometers : rather simple and cheap! employ absorption or interference filters for limiting  ex and  em Spectrofluorometers : more sophisticated and expensive! allow generation of both excitation and emission spectra, employ two grating monochromators