Can you explain the concept of Spectrofluorimetry?
Spectrofluorimetry is a highly sensitive analytical method for the detection and determination of fluorescent compounds at ng or lower level. It has extensively been applied to the analysis of multicomponent systems and some applications in the field of pharmaceutical and clinical analysis are presented as follows.
What is the purpose of a spectrofluorometer?
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.
What is the principle of fluorimetry?
Principle of Fluorimetry: When molecules are irradiated with light of the appropriate frequency, it will be absorbed in about 10-15 seconds. In the process of absorption, the molecules may move from ground to the first excited singlet electronic state.
Why is Spectrofluorometry potentially more sensitive?
is a process by which radiation is produced as a result of a chemical reaction. Why is spectrofluorometry potentially more sensitive than spectrophotometry? For spectrofluorometry, the analytical signal F is proportional to the source intensity P0 and the transducer sensitivity.
How does fluorescence spectroscopy work?
Fluorescence spectroscopy uses a beam of light that excites the electrons in molecules of certain compounds, and causes them to emit light. That light is directed towards a filter and onto a detector for measurement and identification of the molecule or changes in the molecule.
What is fluorimetry and Phosphorimetry?
Fluorimetry and phosphorimetry are analytical techniques based on the absorption of electromagnetic radiation by the processed sample, which contains or generates radiation-absorbing molecules in excited states; when the excited molecules return to the ground state, radiation is emitted, i.e., the excited molecules …
What are primary and secondary filter in fluorimetry?
Filters. There are two filters for the fluorometer: The primary filter or excitation filter or incident light filter isolates the wavelength that will cause the compound to fluoresce (the incident light). The secondary filter isolates the desired emitted light (fluorescent light).
Why is spectrofluorometry potentially more sensitive than UV VIS absorption spectrophotometry?
The reason why fluorescence is more sensitive than UV-Vis absorption is that they are measured in different ways. Absorbance is measured as the difference in intensity between light passing through the reference and the sample, whereas fluorescence is measured directly without any reference beam.
What is fluorescence excitation and emission?
A fluorescence excitation spectrum is when the emission wavelength is fixed and the excitation monochromator wavelength is scanned. In this way, the spectrum gives information about the wavelengths at which a sample will absorb so as to emit at the single emission wavelength chosen for observation.