Fluorescence spectrometry is a fast, simple and inexpensive method to determine the concentration of an analyte in solution based on its fluorescent properties used for measuring compounds in solution.
Therefore in fluorescence spectroscopy, a beam of light with a wavelength varying between 180 and ∼800 nm passes through a solution present in a cuvette. In other words It is a measure from an angle, and the light emitted by the sample.
For instance , in fluorescence spectrometry both an excitation spectrum (the light absorbed by the sample solution) or in other words an emission spectrum (the light emitted by the sample solution) is measured or estimated.
The concentration of the analysis sample is directly proportional with the intensity of the emission.
However,there are several parameters influencing the intensity and shape of the spectrum, record of emission spectrum, the intensity is dependent on the:
- Excitation wavelength of light
- Concentration of the solvent
- Path length of the cuvette
- self-absorption of the sample
Additionally, Fluorescence analysis is suitable for analytes that dissolve in solvents like water, ethanol and hexane.
Above all the analytical sample needs to absorb UV or visible light. After that the analytical sample needs to emit visible or near infra-red radiation.
Therefore,with fluorescence analysis we can do quantitative measurements of a single analysis in solution, (Or one analyte in solution) provided they do not interfere with each other.
In conclusion, an excitation ( called an absorption spectrum) and an emission spectrum of a fluorescent compound can be created.
The absorption spectrum tells us which wavelengths coming towards it can be absorbed by the solution, with the emission or fluorescence spectrum, and after that we can find out which wavelengths are emitted after absorbing the incoming light.
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