High-performance size-exclusion chromatography of humic substances on the hydroxyethyl methacrylate column

HEMA, the copolymer of 2-hydroxyethyl methacrylate and ethylene dimethacrylate, has proven to be a suitable stationary phase for the size-exclusion chromatographic (SEC) characterization of humic substances in a wide pH range. The SEC measurements were performed typically with alkaline borate-based mobile phase: 0.025 mol/l Na2B4O7, 0.025 mol/l NaCl, 0.001 mol/l Na4P2O7, pH 9.1. After calibrating with globular protein standards, apparent molecular weights of typical peat- and coal-derived humic acids were estimated. The average molecular weights determined with the aid of the HEMA column were quite comparable with the values obtained previously by the method of low-pressure gel-permeation chromatography on the Sephadex column.