Determination of phosphate in soil extracts in the field: A green chemistry enzymatic method

摘要

class="kwd-title">Abbreviations: HEPES, 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid; IMAC, immobilized metal-ion affinity chromatography; MESG, 2-amino-6-mercapto-7-methylpurine ribonucleoside; PNP, purine nucleoside phosphorylase (EC 2.4.2.1) class="kwd-title">Method name: Soil phosphate determination in the field class="kwd-title">Keywords: Soil phosphate, Soil phosphorus, Field, Enzymatic, Green chemistry, Open-source portable photometer class="head no_bottom_margin" id="idm140361605876464title">AbstractMeasurement of ortho-phosphate in soil extracts usually involves sending dried samples of soil to a laboratory for analysis and waiting several weeks for the results. Phosphate determination methods often involve use of strong acids, heavy metals, and organic dyes. To overcome limitations of this approach, we have developed a phosphate determination method which can be carried out in the field to obtain results on the spot.This new method uses: class="first-line-outdent">

• Small volumes.

• An enzymatic reaction.

• Green chemistry.First, the soil sample is extracted with deionized water and filtered. Next, an aliquot of the soil extract (0.5 mL) is transferred to a disposable cuvette, containing 0.5 mL of reaction mixture [200 mM HEPES, pH 7.6, 20 mM MgCl2, with 80 nmol 2-amino-6-mercapto-7-methylpurine ribonucleoside (MESG) and 1 unit of recombinant purine nucleoside phosphorylase (PNP; EC 2.4.2.1)], mixed, and incubated for 10 min at field temperature. Absorbance of the completed reaction is measured at 360 nm in open-source, portable photometer linked by bluetooth to a smartphone. The phosphate and phosphorus content of the soil is determined by comparison of its absorbance at 360 nm to a previously prepared standard phosphate curve, which is stored in the smartphone app.