title: A comparison of five lipid extraction solvent systems for lipidomic studies of human LDL.
abstract: Lipidome profile of fluids and tissues is a growing field as the role of lipids as signaling molecules is increasingly understood, relying on an effective and representative extraction of the lipids present. A number of solvent systems suitable for lipid extraction are commonly in use, though no comprehensive investigation of their effectiveness across multiple lipid classes has been carried out. To address this, human LDL from normolipidemic volunteers was used to evaluate five different solvent extraction protocols [Folch, Bligh and Dyer, acidified Bligh and Dyer, methanol (MeOH)-tert-butyl methyl ether (TBME), and hexane-isopropanol] and the extracted lipids were analyzed by LC-MS in a high-resolution instrument equipped with polarity switching. Overall, more than 350 different lipid species from 19 lipid subclasses were identified. Solvent composition had a small effect on the extraction of predominant lipid classes (triacylglycerides, cholesterol esters, and phosphatidylcholines). In contrast, extraction of less abundant lipids (phosphatidylinositols, lyso-lipids, ceramides, and cholesterol sulfates) was greatly influenced by the solvent system used. Overall, the Folch method was most effective for the extraction of a broad range of lipid classes in LDL, although the hexane-isopropanol method was best for apolar lipids and the MeOH-TBME method was suitable for lactosyl ceramides.
authors.affiliation: School of Life and Health Sciences, Aston University, Birmingham, UK. anareis@ua.pt
authors.forename: Ana
authors.initials: A
authors.lastname: Reis
authors.forename: Alisa
authors.initials: A
authors.lastname: Rudnitskaya
authors.forename: Gavin J
authors.initials: GJ
authors.lastname: Blackburn
authors.forename: Norsyahida
authors.initials: N
authors.lastname: Mohd Fauzi
authors.forename: Andrew R
authors.initials: AR
authors.lastname: Pitt
authors.forename: Corinne M
authors.initials: CM
authors.lastname: Spickett
mesh_terms: D000328:Adult; D008078:Cholesterol, LDL / Q000737:chemistry* / Q000302:isolation & purification*; D005260:Female; D064368:Healthy Volunteers; D006801:Humans; D008875:Middle Aged; D012997:Solvents / Q000737:chemistry*
chemical_list: D008078:Cholesterol, LDL; D012997:Solvents
keywords: ANOVA simultaneous component analysis; dual polarity; lipidomics; liquid-liquid extraction; orbitrap; polarity switching