Measurement of Phosphoinositide (PIP) Metabolism via Derivatization-Based Liquid Chromatography-Tandem Mass Spectrometry

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April 29,2025

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The parent ions and the characteristic fragment ions (glycerol backbone with fatty acyl chains) were utilized to distinguish PIP species with distinct fatty acyl chains or identical chains but varying phosphorylation states. Combined with chiral chromatography columns, PIPs sharing the same fatty acyl composition but differing in phosphorylation positions (e.g., PI(3,4)P2 vs. PI(3,5)P2) were effectively separated. This methodology enabled the successful identification of more than 60 PIP species in HeLa cells, demonstrating exceptional selectivity and sensitivity.

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