Capillary electrophoretic behaviour of humic substances in physical gels

We investigated the principles of the capillary electrophoretic behaviour of humic substances (HSs) in physical gels. Long chain (Mr 4000, 6000 and 20,000) polyethylene glycols (PEGs) at concentrations above their entanglement threshold caused the separation of HS fractions according to molecular size differences. Close linear relationships between effective mobilities and mean apparent molecular masses were observed at PEG concentrations between 2.5 and 15%. The efficiency of the separation does not increase in gels of increasing polymer concentrations. The possibility of interactions between HSs and gel-forming polymers was also investigated. Short chain (Mr 400) PEGs, added to the buffer at concentrations from 2.5 to 12.5%, increased the migration times of all HS fractions, but no separation was obtained even at large polymer concentrations, showing that gel formation was essential for the separation. In 2.5% polyvinyl alcohol (PVA) 49 000 all fractions show two unresolved, but well defined peaks. This separation is probably artefactual and depends on the relative concentration of HSs and PVA, as the relative abundance of the peaks changes with the sample concentration.