Rapid and sensitive determination of nucleoside H-phosphonates and inorganice H-phosphonates by high-performance liquid chromatography coupled with flow-injection analysis

Abstract

A coupled high-performance liquid chromatographic (HPLC) and flow-injection analysis (FIA) system was developed for the separation and detection of H-phosphonates. The H-phosphonates are separated by HPLC in the anion-exchange mode and then introduced into an on-line FIA system. In the FIA system, H-phosphonate esters and anhydrides (POP bonds are hydrolysed and oxidized to orthophosphate, the resulting orthophosphate is converted into the heteropoly blue complex and the latter is monitored spectrophotometrically at 830 nm. The system is sensitive and easy to use. The detection limit is 10 −6 M for inorganic, sugar and nucleoside H-phosphonates. The reproducibility of the peak areas was less than 1% (relative standard deviation) for all H-phosphonates.