How to correct absorption spectrum of a fluorescent compound?
- From: "Farooq W" <farooq.w@xxxxxxxxx>
- Date: 27 Jan 2006 07:22:31 -0800
I came across a related problem (of academic interest) in Galen Ewing's
book on Instrumental Analysis.
"The absorption spectrum of a fluorescent substance determined with a
coventional spectrophotometer is likely to be in error because of
fluorescence. Suppose a compound has an absorption maximum (hence an
excitation maximum) at 290 nm and a fluorescence maximum at 350 nm. At
what wavelength would you expect the greatest error. Would the observed
absorbance be too large or too small at this point. Would your answers
be different for a spectrophotometer in which the radiation from the
lamp passes through the cuvet before dispersion in the monochromator"
I think the greatest error would be at 290 nm, where the absorbance
recorded would be too low because "extra" light due to excitation would
be reaching the detector.
For the second spectrophotometer which uses dipersion deivce between
sample and detector one would get the correct spectrum.
Does anyone else has more practical suggestions for the correction of
such spectrum?
.
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