Trapped ion mobility spectrometry and PASEF enable in-depth lipidomics from minimal sample amounts

Area of Science:

• Analytical Chemistry
• Biochemistry
• Mass Spectrometry

Background:

• Comprehensive lipidome characterization is challenging with limited sample volumes.
• Existing methods struggle to identify a wide range of lipids efficiently.

Purpose of the Study:

• To develop a high-sensitivity lipidomics workflow for comprehensive lipidome analysis.
• To improve lipid identification and quantification using advanced mass spectrometry techniques.

Main Methods:

• Utilized nanoflow liquid chromatography coupled with trapped ion mobility spectrometry (TIMS).
• Implemented parallel accumulation-serial fragmentation (PASEF) for enhanced precursor ion analysis.
• Achieved high acquisition speeds (>100 Hz) for rapid MS/MS data generation.

Main Results:

• PASEF increased lipid identification by over threefold compared to standard TIMS-MS/MS in human plasma.
• Attomole sensitivity was achieved, enabling analysis of minute sample volumes (1 µL).
• Compiled a dataset of 1856 lipid collisional cross section (CCS) values from diverse biological samples.

Conclusions:

• Established PASEF as a powerful technique for advanced lipidomics.
• Demonstrated the potential for sensitive, ion mobility-enhanced lipidomics in four dimensions.
• The workflow enables comprehensive lipidome characterization from limited biological material.