Using Rayleigh Scattering to Correct the Inner Filter Effect of the Fluorescence Excitation-Emission Matrix.

Abstract

Excitation-emission matrix (EEM) spectroscopy has been proven to be an effective tool for offline fluorescence analysis. However, the pretreatment of EEM data requires an additional ultraviolet-visible (UV-vis) absorption spectrum for inner filter effect (IFE) correction. This complicates the instrument structure and increases the test flow, thus hindering the practical application of EEM in environmental online monitoring. In this work, Rayleigh scattering in EEM, which is often masked, is leveraged to address this challenge as Rayleigh scattering light itself passes through the sample and experiences absorption. We establish a translation-corrected estimation by the Rayleigh scattering (TCERS) method to estimate absorbance, not only enabling the IFE self-correction of EEM but also providing orthogonal spectroscopy information. TCERS is hierarchically tested in real solutions, simulated turbid liquids, and various natural water samples. Results indicate that the predicted UV-vis absorption spectra have a cosine similarity of over 0.95 with the actual spectra. When using the predicted spectra to correct the IFE of EEM, only about 0.005/1.440 bits of information entropy are lost and the absolute errors in EEM are negligible. The proposed method has the potential to streamline the design of fluorescence spectrometers, making it possible to miniaturize, optimize, and popularize these instruments for various practical applications such as environmental monitoring.