Solid-state CP/MAS ¹³C NMR analysis of particle size and density fractions of a soil incubated with uniformly labeled ¹³C-glucose

Abstract

A soil incubated for 34 days in the absence (control) and presence (treated) of uniformly labelled 13C-glucose was dispersed using an ultrasonic probe and fractionated by sedimentation in water and a polytungstate solution of density 2.0 Mg m3 . The residual substrate carbon was concentrated in the clay and light fractions. Solid state CP/MAS 13C n.m.r. (cross polarization/magic angle spinning 13c nuclear magnetic resonance) spectroscopy was used to characterize the chemical structure of the native soil organic carbon and the residual substrate carbon in the fractions of the control and treated soils. To obtain quantitative results it was essential to determine the spin lattice relaxation times in the rotating frame, T1pH, of the individual carbon types in the spectra as the relaxation behaviour of the native organic materials in the clay fraction was substantially different from that of the residual substrate carbon. After correcting for T1pH effects, a significant linear relationship existed between the signal intensity and 13C content of the samples. This enabled the content, expressed in µmol 13C g-1 fraction, of each type of carbon in the fractions to be calculated. The residual substrate carbon was found to accumulate in predominantly alkyl and O-alkyl structures in both fractions. However, significant amounts of acetal and carboxyl carbon were also observed in the clay fraction. Little if any aromatic or phenolic carbon was synthesized by the soil microorganisms utilizing substrate carbon. Dipolar dephasing CP/MAS 13C n.m.r. experiments were also performed and allowed the proportion of each type of carbon which was protonated and nonprotonated to be estimated. Essentially all of the O-alkyl and acetal carbon, 25-40% of the aromatic carbon and 66-80% of the alkyl carbon was protonated in the fractions isolated from the treated soil