title: An Efficient Single Phase Method for the Extraction of Plasma Lipids.
abstract: 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.
authors.affiliation: Baker IDI, Heart and Diabetes Institute, Melbourne, VIC 3004, Australia. malcolmm@unimelb.edu.au.|King Fahad Medical City, Riyadh 11525, Saudi Arabia. malcolmm@unimelb.edu.au.|The Bio21 Institute of Molecular Science and Biotechnology, University of Melbourne, Melbourne, VIC 3010, Australia. malcolmm@unimelb.edu.au.
authors.forename: Zahir H
authors.initials: ZH
authors.lastname: Alshehry
authors.affiliation: Baker IDI, Heart and Diabetes Institute, Melbourne, VIC 3004, Australia. Christopher.Barlow@bakeridi.edu.au.
authors.forename: Christopher K
authors.initials: CK
authors.lastname: Barlow
authors.affiliation: Baker IDI, Heart and Diabetes Institute, Melbourne, VIC 3004, Australia. jacqui.weir@bakeridi.edu.au.
authors.forename: Jacquelyn M
authors.initials: JM
authors.lastname: Weir
authors.affiliation: Baker IDI, Heart and Diabetes Institute, Melbourne, VIC 3004, Australia. zhouyouping@yahoo.com.
authors.forename: Youping
authors.initials: Y
authors.lastname: Zhou
authors.affiliation: The Bio21 Institute of Molecular Science and Biotechnology, University of Melbourne, Melbourne, VIC 3010, Australia. malcolmm@unimelb.edu.au.
authors.forename: Malcolm J
authors.initials: MJ
authors.lastname: McConville
authors.affiliation: Baker IDI, Heart and Diabetes Institute, Melbourne, VIC 3004, Australia. peter.meikle@bakeridi.edu.au.|King Fahad Medical City, Riyadh 11525, Saudi Arabia. peter.meikle@bakeridi.edu.au.
authors.forename: Peter J
authors.initials: PJ
authors.lastname: Meikle
keywords: 1-butanol/methanol extraction; lipidomics; mass spectrometry