High-performance liquid chromatographic determination of pyrophosphate in the presence of a 20,000-fold excess of orthophosphate.
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.
Yoza
N
N
Department of Chemistry, Faculty of Science, Kyushu University, Fukuoka, Japan.
Akazaki
I
I
Nakazato
T
T
Ueda
N
N
Kodama
H
H
Tateda
A
A
D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D002852:Chromatography, Ion Exchange / Q000379:methods*; D011756:Diphosphates / Q000032:analysis* / Q000302:isolation & purification; D008982:Molybdenum / Q000737:chemistry; D010710:Phosphates / Q000032:analysis* / Q000302:isolation & purification; D012680:Sensitivity and Specificity
D011756:Diphosphates; D010710:Phosphates; D008982:Molybdenum