LC/MS lipid profiling from human serum: a new method for global lipid extraction

Area of Science:

• Biochemistry
• Analytical Chemistry
• Clinical Chemistry

Background:

• Liquid chromatography-mass spectrometry (LC/MS) analyzers have advanced lipidomics research.
• Nine key lipid classes comprise 99% of the human serum lipidome.
• Existing sample preparation methods lack satisfactory recovery for all major lipid classes simultaneously.

Purpose of the Study:

• To develop and validate a novel one-phase extraction (OPE) method for comprehensive human serum lipid profiling.
• To overcome the limitation of incomplete lipid class recovery in current methods.
• To establish a simplified, efficient method for large-scale lipidomics applications.

Main Methods:

• Development and validation of a new one-phase extraction (OPE) method.
• Comparison of the OPE method with established techniques (Folch, Bligh & Dyer) and other solvent systems.
• Incubation of 100 μL human serum with 2 mL of a methanol/chloroform/methyl-tert-butyl ether (MMC) solvent mixture, followed by vortexing and centrifugation.

Main Results:

• The developed MMC one-phase extraction method demonstrated recovery close to 100% for all nine investigated human serum lipid classes.
• The OPE method proved to be simple, rapid, and reproducible.
• The method requires a reduced sample volume (100 μL).

Conclusions:

• The novel MMC one-phase extraction method offers a significant improvement for comprehensive human serum lipid analysis.
• This method simplifies sample preparation, enabling high recovery for all major lipid classes.
• The OPE method facilitates large-scale applications in scientific research and clinical trials for serum lipid profiling.