Abstract
Plant biomass represents a rich nutrient resource for microorganisms. This includes both structural chemical constituents that require degradation and directly accessible soluble compounds such as amino acids. The concentration of free amino acids in plant litter is dynamic and is significantly affected by degradation patterns and leaching. This chapter describes the procedures for the efficient extraction of free amino acids from plant tissues and their subsequent identification and quantification by high pressure liquid chromatography (HPLC). Leaf tissue is ground in liquid nitrogen and extracted in 0.1% HCl. Amino acids in the extract are then allowed to react with 4-nitrophenylisothiocyanate (NPITC) in acetonitrile, and the products are subsequently separated with HPLC on a C-18 reversed-phase column using an acetonitrile-based solvent gradient and UV detection at 340 or 254 nm. Amino acids in peptides and proteins are not measured by this method, unless a digestion step is included. Determining the available pool of free amino acids in plant litter is important to understand the nutrient value of the litter to microorganisms and patterns of litter colonisation and decomposition.
Published Version
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