Assessing Crop Residue Phosphorus Speciation Using Chemical Fractionation and Solution 31P Nuclear Magnetic Resonance Spectroscopy.

At physiological maturity, nutrients in crop residues can be released to the soil where they are incorporated into different labile and non-labile pools. The chemical speciation of phosphorus (P) in crop residues is an important determinant of the fate of this P. In this study, we used chemical fractionation and ³¹P nuclear magnetic resonance (NMR) spectroscopy, first separately and then together, to evaluate the P speciation of mature oat (Avena sativa) residue. Two water extracts (one employing shaking and the other sonication) and two acid extracts (0.2 N perchloric acid and 10% trichloroacetic acid) of these residues contained similar concentrations of orthophosphate (molybdate-reactive P determined by colorimetry) as NaOH-EDTA extracts of whole plant material subsequently analysed by solution ³¹P NMR spectroscopy. However, solution ³¹P NMR analysis of the extracts and residues isolated during the water/acid extractions indicated that this similarity resulted from a fortuitous coincidence as the orthophosphate concentration in the water/acid extracts was increased by the hydrolysis of pyrophosphate and organic P forms while at the same time there was incomplete extraction of orthophosphate. Two methods for isolating and quantifying phospholipid P were also tested, based on solubility in ethanol:ether and ethanol:ether:chloroform. While these methods were selective and appeared to extract only phospholipid P, they did not extract all phospholipid P, as some was detected by NMR in the crop residue after extraction. These results highlight the need for careful interpretation of results from chemical fractionation, as separation can be compromised by incomplete recovery and side reactions.