while conducting the interaction studies between dye and nanoparticles, we observed quenching of dye..how can we confirm the quenching of dye has occurred not because of inner filter effect if both the dye and nanoparticle absorbs the same excitation wavelength?
If the absorbance of the nanoparticles is significant at the excitation and/or emission wavelengths of the fluorophore, then the inner filter effect will contribute to the observed reduction in fluorescence, but it may not be the whole story. There could also be static and/or dynamic quenching. Additionally, light scattering by the nanoparticles could contribute, and there could be FRET between the fluorophore (donor) and nanoparticles (acceptor) if they are very close together.
If you know of another fluorophore that can be excited and emits at similar wavelengths as the first one, but definitely does not interact with the nanoparticles, you can see whether there is any reduction in fluorescence of that fluorophore when the nanoparticles are added. That will allow you to see how much of an inner filter effect/light scattering effect is happening.
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