Electrophoretic evidence of the integrity of humic substances separated by means of electrofocusing

The alkaline extracts of an andic soil, a Bhir horizon of a spodic soil and leonardite which, after electrofocusing, displayed bands in the neutral region of the pH gradient, were fractionated by electrophoresis in phosphate buffer at pH 6.5. The non-migrated fraction gave an electrofocusing profile containing only bands focused at pH values above that of the electrophoretic separation. Electrophoresis in sodium acetate buffer pH 4.8 of humic substances extracted from peat permitted isolation of bands focusing above pH4.8. The distribution of metals after electrophoresis suggests that a great part of humic substances which originate bands above pH 6.5 consists of negatively charged Fe and Al complexes. Addition of EDTA to solutions of humic substances extracted from the samples resulted in the disappearance of bands in the neutral region of the pH gradient in the electrofocusing profile of the Bhir sample, while a consistent lessening of these bands was observed in the leonardite and andic soil samples.