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Benchmarking One-Phase Lipid Extractions for Plasma Lipidomics.

Marcus Höring¹, Christoph Stieglmeier¹, Katja Schnabel¹

¹Institute of Clinical Chemistry and Laboratory Medicine, Regensburg University Hospital, D-93042 Regensburg, Germany.

Analytical Chemistry

|September 1, 2022

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One-phase lipid extraction methods often fail to recover nonpolar lipids like triglycerides and cholesteryl esters from human plasma. Researchers recommend limiting one-phase extractions to polar lipids for accurate lipidomics analysis.

Area of Science:

Lipidomics
Mass Spectrometry
Analytical Chemistry

Background:

Accurate lipidomics analysis relies on efficient lipid extraction, traditionally using two-phase systems.
One-phase extraction methods are increasingly popular due to their ease of use, but their lipid recovery efficiency is not well-established.

Purpose of the Study:

To benchmark the lipid recovery of popular one-phase extraction methods for human plasma.
To evaluate the efficiency of various organic solvents and solvent mixtures for lipid extraction.

Main Methods:

Quantitative flow injection analysis high-resolution mass spectrometry was employed.
The lipid recovery of methanol, ethanol, 2-propanol, 1-butanol, acetonitrile, and binary solvent mixtures was assessed.
Extraction efficiencies for different lipid classes were quantified.

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A Simple Fractionated Extraction Method for the Comprehensive Analysis of Metabolites, Lipids, and Proteins from a Single Sample

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Shotgun Lipidomics of Rodent Tissues

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Published on: November 18, 2022

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Main Results:

Polar lysophospholipids showed sufficient recovery across all tested solvents.
Nonpolar lipids, including triglycerides and cholesteryl esters, had extraction efficiencies below 5% due to precipitation in polar solvents.
Significant amounts of phospholipids precipitated in acetonitrile, indicating substantial lipid loss.

Conclusions:

One-phase extraction methods demonstrate poor recovery of nonpolar lipids from human plasma.
Lipid loss is directly related to solvent polarity and lipid class nonpolarity.
The use of one-phase extractions should be restricted to polar lipid classes unless nonpolar lipid recovery is validated for accurate lipid quantification.