A rapid one-step spectrophotometric assay for orthophosphate that requires a single stable reagent solution is presented. The reagent solution, an aqueous mixture of ammonium molybdate and zinc acetate at pH 5.0, produces a stable complex with orthophosphate that absorbs strongly in the near-visible region of the light spectrum. Response to concentration of phosphate was linear up to 300 microM phosphate with a molar absorptivity of 7200 M-1 cm-1 at 350 nm. The mild conditions for phosphate determination employed in this method are unique, making it particularly suitable for the assay of orthophosphate in the presence of labile organophosphates.
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