A Targeted, Bioinert LC-MS/MS Method for Sensitive, Comprehensive Analysis of Signaling Lipids

Area of Science:

• Lipidomics
• Analytical Chemistry
• Cellular Biology

Background:

• Signaling lipids regulate critical cellular functions.
• Existing methods lack comprehensiveness and face analytical challenges like wide dynamic range and isomeric complexity.
• Comprehensive monitoring of signaling lipids in a single analytical run is currently unavailable.

Purpose of the Study:

• To develop a sensitive and robust targeted liquid chromatography-mass spectrometry (LC-MS/MS) approach for comprehensive signaling lipid analysis.
• To overcome challenges in analyzing diverse signaling lipid classes, including isomeric and isobaric species.
• To enable the simultaneous monitoring of a broad panel of signaling lipids in complex biological samples.

Main Methods:

• A targeted LC-MS/MS workflow was developed.
• Utilized bioinert reversed-phase liquid chromatography coupled with targeted mass spectrometry.
• Employed a simple one-phase sample extraction protocol.

Main Results:

• The method covers 17 different signaling lipid classes and 388 distinct lipid species.
• Achieved high sensitivity and repeatability across various biological matrices.
• Successfully characterized the human plasma signaling lipidome, quantifying 307 endogenous molecular lipid species.
• Investigated signaling lipid alterations during platelet activation, revealing pathway regulations.

Conclusions:

• The presented LC-MS/MS workflow offers a sensitive, robust, and comprehensive solution for signaling lipid analysis.
• Enables unprecedentedly detailed insights into signaling pathways within complex biological systems.
• Facilitates the investigation of lipid signaling in cellular processes and disease states.