Analysis of dilute aqueous multifluorophoric mixtures using excitation-emission matrix fluorescence (EEMF) and total synchronous fluorescence (TSF) spectroscopy: A comparative evaluation

Excitation-emission matrix fluorescence (EEMF) and total synchronous fluorescence (TSF) spectroscopy are two conceptually different fluorescence techniques that have been used to map the fluorescence responses of the fluorophores present in a multifluorophoric mixture. EEMF was introduced four decades back and most of the fluorimeters have the suitable computer program which allows the acquisition EEMF spectra. Recently introduced TSF spectroscopy has been shown to possess good application potential in analytical fluorimetry and has started attracting the attention of analytical chemists. TSF data structure, however, is intrinsically different from EEMF data structure and a better understanding of TSF data structure is crucial to utilising its application potential. In the present work, a comprehensive comparative study between EEMF and TSF spectroscopic data set was performed by taking aqueous mixtures containing low concentrations of benzo[a]pyrene, chrysene, and pyrene as test case. The EEMF and TSF data structures were clearly explained by taking pyrene as an example. The effects of Rayleigh and Raman scattering on the quality of EEMF and TSF data sets were studied. EEMF and TSF data sets of dilute aqueous mixtures of benzo[a]pyrene, chrysene, and pyrene were subjected to three chemometric techniques PARAFAC, N-PLS, and MCR-ALS analysis. TSF data set in particular was found to be highly attuned to MCR-ALS analysis. Obtained results of chemometric analyses on EEMF and TSF data sets show that TSF data of dilute aqueous mixtures provides more accurate spectral and concentration information than EEMF data sets. Therefore, TSF spectroscopy could be considered as an alternate to the EEMF for the analyses of dilute multifluorophoric mixtures.