High-performance liquid chromatographic determination of pyrophosphate in the presence of a 20,000-fold excess of orthophosphate

✍ Scribed by Norimasa Yoza; Izumi Akazaki; Tetsuya Nakazato; Nobuyuki Ueda; Hiroki Kodama; Akira Tateda

• Book ID: 102986402
• Publisher: Elsevier Science
• Year: 1991
• Tongue: English
• Weight: 616 KB
• Volume: 199
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(91)90102-y

✦ Synopsis

An HPLC method was based on anion-exchange separation of pyrophosphate (diphosphate) and orthophosphate and postcolumn spectrophotometric detection at 140 degrees C with a molybdenum(V)-molybdenum(VI) reagent. The reagent was easy to prepare, stable for at least 6 months at room temperature, and ready for the determination of pyrophosphate and orthophosphate by the so-called heteropoly blue method without use of any reducing agent. A photodiode-array detector for HPLC indicated the spectral characteristics of the heteropoply blue complex that was detectable at 330-800 nm. The HPLC method had a wide dynamic range from 3 x 10(-7) to 5 x 10(-4) M for both pyrophosphate and orthophosphate with a relative standard deviation of measurement of 10 approximately 2%. Pyrophosphate of 5 x 10(-7) and 5 x 10(-6) M, respectively, could be determined in the presence of a 20,000-fold excess of orthophosphate; 0.01 and 0.1 M.