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Updated: Aug 10, 2026

High-Throughput Measurement and Classification of Organic P in Environmental Samples
Published on: June 8, 2011
Enzymatic determination of phosphate in conjunction with high-performance liquid chromatography
Abstract:
A selective assay for orthophosphate in complex matrices was developed based on the nucleoside phosphorylase catalyzed conversion of inosine and orthophosphate of hypoxanthine. The enzyme reaction was using only 0.28 units/assay was allowed to proceed for 30 min before quenching. Separation of inosine and hypoxanthine was performed by reversed-phase high-performance liquid chromatography. Quantitation of the hypoxanthine peak was found to be linear with orthophosphate up to 30 micrograms/g. A detection limit of 0.75 ppm could be obtained after dialysis of the commercial enzyme. Interference studies showed that the enzymatic assay unlike the colorimetric molybdate-blue technique was essentially unaffected by complex matrices such as serum, urine, polyphosphates, and phosphoesters.
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