UHPLC-TIMS-PASEF^®-MS for Lipidomics: From Theory to Practice

Area of Science:

• Analytical Chemistry
• Biochemistry
• Molecular Biology

Background:

• Lipidomics analysis faces challenges in identifying isomeric and structurally similar lipids.
• Traditional mass spectrometry methods can suffer from co-elution and ion coalescence, impacting data quality.
• High-throughput lipidomic studies require rapid and accurate analytical techniques.

Purpose of the Study:

• To present a data-dependent Trapped Ion Mobility Spectrometry (TIMS) workflow utilizing Parallel Accumulation Serial Fragmentation (PASEF) for lipidomics.
• To highlight the advantages of TIMS-PASEF in enhancing lipid identification and structural characterization.
• To demonstrate the suitability of this workflow for high-throughput lipidomics.

Main Methods:

• Implementation of a data-dependent TIMS-PASEF acquisition strategy.
• Utilizing the additional separation dimension provided by TIMS for ion mobility.
• Employing PASEF for rapid, parallel ion accumulation and serial fragmentation.

Main Results:

• TIMS provides accurate collision cross-section (CCS) measurements, aiding in distinguishing lipid isomers.
• PASEF enables fast data acquisition, significantly increasing analytical throughput.
• The combination of TIMS and PASEF reduces spectral complexity by minimizing co-elution and ion coalescence.
• Improved data quality and increased confidence in lipid identifications were achieved.

Conclusions:

• The TIMS-PASEF workflow offers a powerful approach for advanced lipidomics analysis.
• This technique enhances the structural elucidation capabilities and efficiency of mass spectrometry-based lipidomics.
• The presented method is well-suited for demanding high-throughput lipidomic applications.