Determination of concentrations of spectrally overlapping compounds has special difficulties. Several methods are available to calculate the constituents' concentrations in moderately complex mixtures. A method which can provide information about spectrally hidden components in mixtures is very useful. Two methods powerful in resolving spectral components are compared in this paper. The first method tested is the Derivative Matrix Isopotential Synchronous Fluorimetry (DMISF). It is based on derivative analysis of MISF spectra, which are constructed using isopotential trajectories in the Excitation-Emission Matrix (EEM) of background solution. For DMISF method, a mathematical routine fitting the 3D data of EEMs was developed. The other method tested uses classical Least Squares Fitting (LSF) algorithm, wherein Rayleigh- and Raman-scattering bands may lead to complications. Both methods give excellent sensitivity and have advantages against each other. Detection limits of DMISF and LSF have been determined at very different concentration and noise levels.
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