3. 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
4. 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.
5.
6. 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.
12. 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
