Analysis of inositol mono- and polyphosphates by gas chromatography/mass spectrometry and fast atom bombardment

Abstract

The electron ionization spectra of all of the positional isomers of myo-inositol monophosphate and of myo-inositol 1,2-cyclic phosphate were obtained by gas chromatography/mass spectrometry of the pertrimethylsilyl derivatives. The fragmentation pattern of pertrimethylsilyl myo-inositol-1-phosphate was studied using deuterium labeling. The phosphate moiety was found to direct fragmentation to produce fragment ions of useful intensity with specific carbon retention. The spectrum of pertrimethylsilyl myo-inositol-1,4-bisphosphate is also described. An electron impact gas chromatographic/mass spectrometric method for myo-inositol-1-phosphate has been developed, which has a sensitivity to a level of 0.1 pmol. The positive and negative ion fast atom bombardment spectra of myo-inositol hexakis(disodium phosphate) and myo-inositol hexakis(dihydrogen phosphate) are described. The lesser-phosphorylated inositol polyphosphates were also studied, including inositol pentakis and inositol tetrakis(dihydrogen phosphates) as well as D-myo-inositol-1,4,5-trisphosphate and D-myo-inositol-1,4-bisphosphate from human red blood cells. The sensitivity of fast atom bombardment for the measurement of the latter two substances allows their detection to a level of about 10 nmol. The fast atom bombardment spectrum of synthetic myo-inositol 1,2-cyclic phosphate revealed variable amounts of a dimer produced during its preparation.