Overestimation of the importance of phytate in NaOH-EDTA soil extracts as assessed by (31)P NMR analyses

Abstract

Solution 31P NMR analysis of NaOH-EDTA extracts is the most widely used method for determining the speciation of organic P in soils. In this paper, we bring together results from a number of our recent studies that highlight some of the difficulties in obtaining accurate quantitative results and propose that these difficulties may have resulted in the systematic overestimation of soil phytate concentrations. The first problem is peak identification, which is complicated by variations in chemical shift with pH and ionic strength, and the impracticality of the primary 31P chemical shift reference (85% H3PO4). Unambiguous peak assignment can be achieved by spiking suspected compounds directly into soil extracts prepared for NMR analysis; this ensures native and added compounds experience exactly the same chemical environment. The second problem is quantifying NMR signal for individual species in the crowded and overlapping phosphate monoester region. Spectral deconvolution has been employed for this purpose, but very different results are obtained depending on whether or not the fitting procedure includes a broad signal that probably comes from P in large, " humic" compounds. In particular, failing to consider this broad peak in the deconvolution procedure results in overestimation of small organic P compounds, including phytate. The ready decomposition of phytate is demonstrated with an incubation experiment in which phytate added at various concentrations to a calcareous soil is shown to be decomposed to produce orthophosphate. The decomposition follows a first order exponential decay with a half-life of 4-8weeks. © 2011 Elsevier Ltd.