Analysis of Inositol Phosphates, Diphosphoinositol Phosphates, and Their Analogs by HPLC on an Acid-Compatible Stationary Phase with Detection by Complexation with Ferric Ion.
Whitfield HL., Sprigg C., Riley AM., Shipton ML., Potter BVL., Brearley CA.
High-performance anion-exchange chromatography with acid eluents allows for effective separation of myo-inositol tetrakisphosphates, myo-inositol pentakisphosphates, diphospho-myo-inositol phosphates (inositol pyrophosphates), inositol hexakisphosphates, inositol pyrophosphate analogs, and carbohydrate-based analogs of inositol phosphates. All are detectable by postcolumn addition of ferric ion. In the following, we provide examples drawn from characterization of ITPK, IPK1, and IP3 3K enzymes.