The present study proposes an approach to use the derived
absorbance
spectral parameter (D
abs) as a tool for sensitive fluorescence measurement
of optically dense multifluorophoric systems. Mishra and group have
demonstrated the use of D
abs in obtaining
optimum offset wavelength for sensitive fluorescence measurement in
synchronous fluorescence spectra (SFS) (Anal. Chim. Acta
2016, 940, 113–119; Anal. Chim. Acta
2008, 630 (1), 47–56). The D
abs is obtained
with the knowledge of UV–vis absorbance spectral data of the
system under study and fluorescence observation filed (FOF) parameters
of the instrument. D
abs is proportional
to the observed fluorescence intensity of the system. Excitation of
an optically dense sample at D
abs maximum
leads to maximum fluorescence intensity obtainable for that system
even in the presence of considerable inner filter effect (IFE). The
earlier studies were applicable to SFS at right-angle fluorescence
measurement geometry and for samples showing only primary IFE. In
the present study, the work has been expanded to general fluorescence
measurements, even when both primary and secondary IFE is present
and for different fluorescence measurement geometries (right-angle,
45° and front-face). The procedure for obtaining D
abs is simple and can be applied to other analyte-specific/unconventional
fluorescence measurement strategies reported in the literature.