Evidence from surface tension and fluorescence data of a pyrene-assisted micelle-like assemblage of humic substances.

Surface activity and fluorescence of humic substances (HS) and HS/pyrene solutions were monitored under various pH conditions. For HS alone the surface tension of the solutions decreased with increasing acidity, with a minimum at around pH 4. This effect, which is a consequence of an increase in the amphiphilic character of structures, is much more pronounced in humic (HA) than in fulvic acids (FA). The addition of pyrene (0.1 micromolL(-1)) results, for HA, in a marked reduction in the migration of amphiphilic species to the solution surface. FA profiles are not modified in presence of pyrene at that concentration. A decrease in the pyrene I1/I3 ratio in HS solutions shows that below pH 9 pyrene molecules react progressively to the change to a more hydrophobic environment, the greatest effect being observed at around pH 6 to 7. These signals are followed by a significant increase in the pyrene excimer fluorescence (lambda(exc)/lambda(em)=334 nm/450 nm), which is a consequence of the proximity of pyrene molecules. For FA, the I1/I3 decrease is less significant and no excimers develop. This set of effects is explained in view of conformational adjustments of HS, mainly HA, which become arranged in micelle-like domains in aqueous solution, the aromatic moieties being assembled around the pyrene molecules.

Fluorescence fingerprint of fulvic and humic acids from varied origins as viewed by single-scan and excitation/emission matrix techniques.

Excitation/emission matrix (EEM), single-scan excitation and synchronous fluorescence spectra of a series of FA and HA from distinct environments are presented. The EEM plots show at least four spectral features whose corresponding Ex/Em pairs relate to the alpha', alpha, beta and gamma (or delta) fluorophores previously found in natural waters spectra. The alpha' and alpha peaks, which identify typical humic-like components, are present in all samples, independently of the organic matter (OM) source. In FA, their Ex/Em pairs are approximately 260 nm/460 nm and approximately 310 nm/440 nm, respectively. In HA their excitation and emission maxima are red-shifted, the corresponding Ex/Em pairs being located at approximately 265 nm/525 nm and approximately 360 nm/520 nm, respectively. The appearance of beta and gamma (or delta) peaks is dependent both on the OM origin and on HS aging. The former (Ex/Em approximately 320 nm/430 nm), that has been associated with the incidence of marine humic-like material, is present only in a few marine and estuarine HA. It emerges as a shoulder on the alpha peak and its detection is dependent on a balance between its magnitude and the magnitude and emission maxima location of the alpha peak. The gamma (or delta) peak (Ex/Em approximately 275 nm/315 nm in FA, and approximately 275 nm/330 nm in HA), on the other hand, is better visualized in FA than in HA diagrams. It has typical protein-, mainly tryptophan-like, fluorescence properties and appears with varied significance in a few marine and estuarine samples being hardly detected in samples from exclusively terrestrial environments. It is also shown in this study that with selected lambda(ex), lambda(em) and (delta)(lambda) values, regular emission, excitation and synchronous spectra can, together, provide a good picture of the OM sources and aging for extracted HS.