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An Efficient Single Phase Method for the Extraction of Plasma Lipids

Baker IDI, Heart and Diabetes Institute, Melbourne, VIC 3004, Australia
King Fahad Medical City, Riyadh 11525, Saudi Arabia
The Bio21 Institute of Molecular Science and Biotechnology, University of Melbourne, Melbourne, VIC 3010, Australia
Author to whom correspondence should be addressed.
Academic Editor: Per Bruheim
Metabolites 2015, 5(2), 389-403;
Received: 17 April 2015 / Revised: 4 June 2015 / Accepted: 8 June 2015 / Published: 17 June 2015
(This article belongs to the Special Issue Metabolomic Methodology)
PDF [341 KB, uploaded 18 June 2015]


Lipidomic approaches are now widely used to investigate the relationship between lipid metabolism, health and disease. Large-scale lipidomics studies typically aim to quantify hundreds to thousands of lipid molecular species in a large number of samples. Consequently, high throughput methodology that can efficiently extract a wide range of lipids from biological samples is required. Current methods often rely on extraction in chloroform:methanol with or without two phase partitioning or other solvents, which are often incompatible with liquid chromatography electrospray ionization-tandem mass spectrometry (LC ESI-MS/MS). Here, we present a fast, simple extraction method that is suitable for high throughput LC ESI-MS/MS. Plasma (10 μL) was mixed with 100 μL 1-butanol:methanol (1:1 v/v) containing internal standards resulting in efficient extraction of all major lipid classes (including sterols, glycerolipids, glycerophospholipids and sphingolipids). Lipids were quantified using positive-ion mode LC ESI-MS/MS. The method showed high recovery (>90%) and reproducibility (%CV < 20%). It showed a strong correlation of all lipid measures with an established chloroform:methanol extraction method (R2 = 0.976). This method uses non-halogenated solvents, requires no drying or reconstitution steps and is suitable for large-scale LC ESI-MS/MS-based lipidomic analyses in research and clinical laboratories. View Full-Text
Keywords: lipidomics; 1-butanol/methanol extraction; mass spectrometry lipidomics; 1-butanol/methanol extraction; mass spectrometry

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Alshehry, Z.H.; Barlow, C.K.; Weir, J.M.; Zhou, Y.; McConville, M.J.; Meikle, P.J. An Efficient Single Phase Method for the Extraction of Plasma Lipids. Metabolites 2015, 5, 389-403.

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