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Tandem size exclusion chromatography-Polyacrylamide gel electrophoresis of humic acids
Authors:Oleg A Trubetskoj  Olga E Trubetskaya  Gaida V Afanasieva  Olga I Reznikova  Bernardo Hermosin  Cesareo Saiz-Jimenez
Abstract:Humic acids (HAs) from four soils were fractionated by size exclusion chromatography (SEC) on Sephadex G-75. Three fractions were obtained in all humic acids, collected and assayed by Polyacrylamide gel electrophoresis (PAGE). The unfractionated HA from each soil was used as reference. Each chromatographic fraction formed one electrophoretic zone corresponding closely to one band of the reference sample with some admixture of the fractions preceding or following. The results indicate that fractionation of HAs by tandem SEC-PAGE can be successfully used for obtaining fractions of reduced polydispersity and different electrophoretic mobilities. Pyrolysis/methylation-gas chromatography-mass spectrometry of the full size preparations of HA and fractions with exactly defined molecular size and electrophoretic mobility showed a different distribution in humic components, mainly lipids, lignin derivatives and N-containing compounds.
Keywords:humic acid fractionation  size exclusion chromatography  gel electrophoresis  pyrolysis  methylation-gas chromatography-mass spectrometry  lipids  lignin derivatives
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