Supercritical fluid chromatography-mass spectrometry enables simultaneous measurement of all phosphoinositide regioisomers

Phosphoinositide species, differing in phosphorylation at hydroxyls of the inositol head group, play roles in various cellular events. Despite the importance of phosphoinositides, simultaneous quantification of individual phosphoinositide species is difficult using conventional methods. Here we developed a supercritical fluid chromatography-mass spectrometry method that can quantify the molecular species of all seven phosphoinositide regioisomers. We used this method to analyze (1) profiles of phosphoinositide species in mouse tissues, (2) the effect of lysophosphatidylinositol acyltransferase 1-depletion on phosphoinositide acylchain composition in cultured cells, and (3) the molecular species of phosphatidylinositol-3-phosphate produced during the induction of autophagy. Although further improvement is needed for the absolute quantification of minor phosphoinositide regioisomers in biological samples, our method should clarify the physiological and pathological roles of phosphoinositide regioisomers at the molecular species level.

Automated summary

Phosphoinositide species, with different phosphorylation patterns, participate in various cellular events. A newly developed chromatography-mass spectrometry method allows for quantification of all seven phosphoinositide regioisomers, facilitating research on their physiological and pathological roles. Further improvements are needed for absolute quantification of minor phosphoinositide regioisomers in biological samples.