Protocol for untargeted lipidomics of human serum using LC-TIMS-PASEF

Area of Science:

• Biochemistry
• Analytical Chemistry
• Metabolomics

Background:

• Lipidomics is crucial for understanding biological processes.
• Existing methods for serum lipidomics can be resource-intensive.
• A need exists for efficient and comprehensive human serum lipid profiling.

Purpose of the Study:

• To present a detailed protocol for untargeted lipidomics of human serum.
• To enable comprehensive lipid profiling using minimal sample volume.
• To facilitate integration with other omics data.

Main Methods:

• Protocol development for project design and sample preparation.
• Implementation of reversed-phase liquid chromatography coupled with trapped ion mobility spectrometry and parallel accumulation serial fragmentation (LC-TIMS-PASEF).
• Detailed procedures for quality control and data processing.

Main Results:

• A robust protocol for untargeted lipidomics of human serum.
• Comprehensive lipid profiling from only 5 μL of serum.
• Annotation of lipids across major classes including glycerophospholipids, glycerolipids, sphingolipids, sterols, and fatty acyls.

Conclusions:

• The developed protocol offers an efficient approach for human serum lipidomics.
• The protocol generates high-quality data suitable for statistical analysis.
• This method supports multi-omics integration for deeper biological insights.