Micelle-like conformation of humic substances as revealed by size exclusion chromatography

To gain further insight into the macromolecular behaviour of humic substances we treated a humic material with simple organic compounds and followed the change in molecular size distribution. Monocarboxylic, dicarboxylic, and tricarboxylic acids shifted the humic absorbance from high to low molecular sizes in size exclusion chromatograms. Mineral acids, phenol, alcohols, were not able to produce the same shift and gave total absorbance at the column void volume as in the case of humic substances alone. Our results are evidence of the micellar behaviour of humic substances in solution and of the importance of hydrophobic bondings in holding humic molecules together. The organic acids enter in the interior of the humic micelle and alter its stereochemical hydrophobic arrangement. In alkaline conditions, the developed negative charges disrupt the apparent high molecular size configuration and disperse the humic material into small-size micelles. This macromolecular property, that we report for the first time, may be of great importance in understanding the biological activity and the overall environmental behaviour of humic substances.