Gradient ion chromatography of inositol phosphates

Abstract

Inositol phosphates including phytic acid were separated in 30 min by gradient ion chromatography with postcolumn derivatization. All four pentakisphosphates were resolved, while four tetrakisphosphate peaks were detected. The limits of detection for all polyphosphates, including tris- and bisphosphates, were between 1 and 2 nmol. The method was used to compare nonenzymatic dephosphorylation of inositol hexakisphosphate at pH 4.0 versus pH 10.8. The only pentakisphosphate detected in calf brains was identified as myo-inositol 1,3,4,5,6-pentakisphosphate. The major pentakisphosphate in raw soybean seeds was myo-inositol 1,2,4,5,6-pentakisphosphate of unknown enantiomeric composition, while lesser amounts of myo-inositol 1,2,3,4,5-pentakisphosphate of unknown enantiomeric composition, myo-inositol 1,2,3,4,6-pentakisphosphate, and myo-inositol 1,3,4,5,6-pentakisphosphate were also present.