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The composition and molecular residence time of soil organic matter (SOM) in four particle-size fractions (POM >200 microm, POM 63-200 microm, silt and clay) were determined using Curie-point pyrolysis/gas chromatography coupled on-line to mass spectrometry. The fractions were isolated from soils, either continuously with a C(3) wheat (soil (13)C value = -26.4 per thousand), or transferred to a C(4) maize (soil (13)C value = -20.2 per thousand) cropping system 23 years ago. Pyrograms contained up to 45 different pyrolysis peaks; 37 (ca. 85%) were identifiable compounds. Lignins and carbohydrates dominated the POM fractions, proteins were abundant, but lignin was (nearly) absent in the silt and clay fractions. The mean turnover time (MRT) for the pyrolysis products in particulate organic matter (POM) was generally <15 years (fast C pool) and 20-300 years (medium or slow C pools) in silt and clay fractions. Methylcyclopentenone (carbohydrate) in the clay fraction and benzene (mixed source) in the silt fraction exhibited the longest MRTs, 297 and 159 years, respectively. Plant-derived organic matter was not stored in soils, but was transformed to microbial remains, mainly in the form of carbohydrates and proteins and held in soil by organo-mineral interactions. Selective preservation of plant-derived OM (i.e. lignin) based on chemical recalcitrance was not observed in these arable soils. Association/presence of C with silt or clays in soils clearly increased MRT values, but in an as yet unresolved manner (i.e. 'truly' stabilized, or potentially still 'labile' but just not accessible C).

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Summary Recent studies have pointed to the occurrence in soil organic matter of an insoluble macromolecular fraction, resistant to drastic alkali and acid hydrolysis. This non‐hydrolysable fraction may contribute to the stable carbon pool in the soil and thus be important for the global carbon budget. We have developed a method to isolate such chemically resistant components, whilst ensuring complete elimination of the hydrolysable constituents of the organic matter but avoiding the formation of insoluble compounds via Maillard‐type condensation reactions. Maize leaves, material especially susceptible to artefact formation, were used for this optimization. Several of the treatments that we tested, including the Klason lignin protocol, proved unsuitable. The most suitable protocol, by progressive hydrolysis with trifluoroacetic and hydrochloric acid, revealed a non‐hydrolysable fraction in maize leaves accounting for about 5% by weight of the leaves and corresponding chiefly to lignin and condensed tannins. The protocol was applied to a forest soil and to the soil from an adjacent plot cleared 35 years ago and since cropped continuously with maize. The abundance, chemical composition and sources of the non‐hydrolysable fraction of these two soils were determined by a combination of spectroscopy, pyrolysis and electron microscopy. This fraction accounted for about 6% of the total organic carbon of both soils; it contains aliphatic moieties, black carbon, melanoidins and, we think, condensed tannins.

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We isolated the non-hydrolysable macromolecular organic fraction (insoluble fraction resistant to drastic laboratory hydrolyses) from a temperate, loamy, forest soil (Lacade´e, France) and from the soil of an adjacent plot cleared 35 years ago and continuously cropped with maize. The quantitative, morphological (light, scanning and transmission electron microscopy) and isotopic (bulk 13 C, individual compound 13 C and radiocarbon dating) features of these two non-hydrolysable fractions were determined and compared. It appeared that: (i) extensive degradation of the non-hydrolysable material inherited from the forest soil occurred upon cropping, revealing that its resistance to drastic laboratory hydrolyses is not paralleled by a great resistance to natural biodegradation triggered by change in land use; (ii) only small inputs of maize-derived compounds occurred in the non-hydrolysable fraction of the cultivated soil so that, in spite of extensive degradation, the forest-inherited carbon still predominates; (iii) the nonhydrolysable fractions of both soils comprise the same components (wood debris, spores, pollen, and, predominantly, granular organic aggregates), which correlate with the previously identified chemical components (charcoal, aliphatic lipid components and melanoidin-like components); (iv) the non-hydrolysable fraction of the cropped soil shows a greater contribution of aliphatic moieties, reflecting differential degradation of the components of the non-hydrolysable material inherited from the forest soil; (v) this degradation resulted in enrichment in the oldest components; and (vi) no relationship is observed, in the two Lacade´e soils, between resistance to drastic laboratory hydrolyses, on the one hand, and stability towards biodegradation in situ, on the other. These observations, added to recent ones on other types of soils, suggest that such a conspicuous uncoupling between non-hydrolysable and stable carbon is probably a general feature of organic matter in soil as opposed to sedimentary organic matter.