Two-dimensional NMR studies of size fractionated Suwannee River Fulvic and Humic Acid Reference

Two-dimensional phase sensitive C-13,H-1 correlation spectra were applied to the investigation of substructures in size fractions obtained by tangential flow multistage ultrafiltration (MST-UF) of humic substances (HS) Suwannee River Fulvic Acid Reference (HS SR FA) and Suwannee River Humic Acid Reference (HS SR HA), purchased from the International Humic Substances Society (IHSS). After size fractionation with MST-UF the HS samples give well resolved two-dimensional C-13,H-1-correlated NMR spectra which offer a great potential for substructure elucidation and even quantification. It is shown that low molecular size lignin moieties undergo demethylation of the methoxy groups, accompanied by removal of the phenylpropane side chains and subsequent reaggregation of the aromatic rings. These findings provide insight into the processes of lignin degradation. Only the fraction > 100 kDa contains macromolecules that have spin-spin relaxation times too short for investigations employing NMR multipulse sequences.