Fluorescence quenching of polycyclic aromatic compounds by humic substances. Part 1. Methodology for the determination of sorption coefficients

An introduction to the methodology of fluorescence quenching measurements a s an experimental tool for investigating the sorption of hydrophobic organi c substances by dissolved organic matter (OM) is given. Special attention i s paid to the implications of inner-filter effects (IFEs) on the determinat ion of sorption coefficients. The essential aspects of the theoretical desc ription of IFEs are outlined and Various procedures for the IFE correction of fluorescence quenching measurements in the presence of dissolved OM are presented. Geometric parameters relevant for the quantitative description o f IFE applying a 90 degrees arrangement between fluorescence excitation and detection are determined by different methods, newly developed or adapted from the literature, for the fluorescence spectrometer used in this work. M oreover, an experimental validation of the applicability of different IFE c orrection procedures is performed. In order to evaluate the variations of s orption coefficients that result from the use of different IFE correction p rocedures, the quenching of the fluorescence of the well-known polycyclic a romatic compound pyrene by a commercially available humic substance in aque ous solution is investigated. Compared to the apparent sorption coefficient , which is obtained without any IFE correction, the sorption coefficients d etermined with the correction of primary IFE or with the correction of prim ary and secondary IFE are on average ca. 40 or 60% lower. The application o f different IFE correction procedures allows the determination of sorption coefficients with appropriate precision of +/-15%.