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DETERMINATION OF TOTAL CARBOHYDRATES

S. Mansfield
Published 2005 · Chemistry

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Plant cell walls are an intricate composite matrix consisting largely of lignin, a phenolic-derived polymer, and the structural carbohydrates, primarily cellulose and hemicelluloses. In addition, minor amounts of pectin and protein are also present, but only represent a small fraction to the secondary xylem of plants, which forms the bulk of terrestrial plant biomass and provides a substrate for litter-degrading microorganisms. Several approaches have been developed to quantify cell wall constituents. This chapter describes two analytical procedures to accurately measure total cell wall carbohydrates. Plant litter is ground in liquid nitrogen and hydrolysed in 72% H2SO4. This secondary acid hydrolysis converts the polymeric carbohydrates to their monomeric fractions, which can be separated, identified and quantified by high performance anion exchange liquid chromatography (HPAELC) with water as the mobile phase and electrochemical or refractive index detection. Alternatively, the monomers are derivatised by acetylation and separated, identified and quantified by gas chromatography (GC) with helium as carrier gas and flame ionisation detection (FID). Both chromatographic methods have been successfully applied to various types of litter originating from diverse environments and representing different degrees of decomposition.
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