Molecular properties of the Humeome of two calcareous grassland soils as revealed by GC/qTOF-MS and NMR spectroscopy

Abstract

A Humeomic fractionation revealed the humus molecular composition of two uncropped calcareous soils of Northern France and differentiated the soils Humeome by extracting humic components first unbound to the organo-mineral matrix and then liberated from their progressively stronger intermolecular and intramolecular ester and ether linkages. We separated organo- (ORG1-3) and water-soluble (AQU2 and AQU4) fractions, a final extractable fraction (RESOM) and soil residues. Organo-soluble fractions were studied by GC coupled with high-resolution mass spectrometry (GC/qTOF-MS), all fractions underwent mono- and two-dimensional liquid-state NMR (except for the iron-rich AQU4 fraction), while solid-state ¹³C-CPMAS-NMR spectroscopy analyzed soil residues. The Calcaric Leptosol (A) showed a larger mass extraction than the Calcaric Cambisol (B), and a greater cumulative C and N content in its Humeome. Both soils showed the greatest weight yield for AQU4 fraction, followed by ORG2, RESOM, ORG1, AQU2, and ORG3. ORG2 was the most differentiating fraction between the two soils for both compound concentration and diversity, showing a larger C content for soil A than for soil B and a different distribution in aromatic compounds, fatty acids, and dicarboxylic acids. No significant differences between soils were found for ORG 3, suggesting similar processes of OM stabilization for its recalcitrant components, mostly hydrophobic esters of alkanoic, hydroxy, and aromatic acids with linear alkanols. We confirmed that Humeomic fractionation coupled to advanced analytical instrumentations enabled a detailed molecular characterization of the soil Humeome and differentiated between the two calcareous grassland soils and the other soils previously subjected to Humeomics.

Keywords: Chemical fractionation; GC/qTOF-MS; Humeomics; Humic molecules; NMR; Soil organic matter.