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Related Experiment Video

Updated: Jan 9, 2026

Quantitative Analysis of the Cellular Lipidome of Saccharomyces Cerevisiae Using Liquid Chromatography Coupled with Tandem Mass Spectrometry
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A sensitive LC-MS/MS method for quantifying phosphatidic acid species using basic mobile phases.

Kotaro Hama1, Shohei Azuma2, Yuko Fujiwara2

  • 1Faculty of Pharmaceutical Sciences, Teikyo University, 2-11-1 Kaga, Itabashi-ku, Tokyo, 173-8605, Japan; Advanced Comprehensive Research Organization (ACRO), Teikyo University, 2-11-1 Kaga, Itabashi-ku, Tokyo, 173-8605, Japan.

Biochemical and Biophysical Research Communications
|December 3, 2025
PubMed
Summary
This summary is machine-generated.

This study presents a new, highly sensitive liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for quantifying phosphatidic acid (PA) molecular species. The developed technique enables accurate analysis of PA

Keywords:
LC-MS/MSPhosphatidic acid

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A Mass Spectrometry-Based Approach to Identify Phosphoprotein Phosphatases and their Interactors
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Area of Science:

  • Lipidomics
  • Cell Biology
  • Analytical Chemistry

Background:

  • Phosphatidic acid (PA) is a vital lipid precursor and signaling molecule.
  • PA's cone shape influences membrane curvature, and its diverse molecular forms necessitate precise quantification.
  • Existing LC-MS/MS methods face challenges due to PA's phosphate group chelating with HPLC columns.

Purpose of the Study:

  • To develop a highly sensitive and straightforward LC-MS/MS method for analyzing phosphatidic acid (PA) molecular species.
  • To overcome limitations in current PA quantification methods.
  • To enable detailed studies on PA metabolism and function.

Main Methods:

  • Utilized a metal-free column to prevent chelation issues.
  • Employed a basic mobile phase for improved separation and detection.
  • Applied liquid chromatography-tandem mass spectrometry (LC-MS/MS) for high-sensitivity analysis.

Main Results:

  • Successfully quantified various PA molecular species with high sensitivity and accuracy.
  • Analyzed PA and other phospholipid species in HEK293 cells, identifying primary fatty acids in PA (C16:0, C18:1).
  • Observed significant increases in PA species in THP-1 cells upon phorbol 12-myristate 13-acetate stimulation.

Conclusions:

  • The developed metal-free LC-MS/MS method provides a sensitive and accurate approach for PA analysis.
  • This method facilitates the investigation of PA's diverse roles in biological systems.
  • The findings support the utility of this method for studying PA metabolism and function in various cell models.