Abstract
Lipid extraction using a monophasic chloroform/methanol/water mixture, coupled with functional group selective derivatization and direct infusion nano-ESI-high-resolution/accurate MS, is shown to facilitate the simultaneous analysis of both highly polar and nonpolar lipids from a single retina lipid extract, including low abundance highly polar ganglioside lipids, nonpolar sphingolipids, and abundant glycerophospholipids. Quantitative comparison showed that the monophasic lipid extraction method yielded similar lipid distributions to those obtained from established "gold standard" biphasic lipid extraction methods known to enrich for either highly polar gangliosides or nonpolar lipids, respectively, with only modest relative ion suppression effects. This improved lipid extraction and analysis strategy therefore enables detailed lipidome analyses of lipid species across a broad range of polarities and abundances, from minimal amounts of biological samples and without need for multiple lipid class-specific extractions or chromatographic separation prior to analysis.
Highlights
Lipid extraction using a monophasic chloroform/ methanol/water mixture, coupled with functional group selective derivatization and direct infusion nano-ESI-highresolution/accurate MS, is shown to facilitate the simultaneous analysis of both highly polar and nonpolar lipids from a single retina lipid extract, including low abundance highly polar ganglioside lipids, nonpolar sphingolipids, and abundant glycerophospholipids
Given that the monophasic retina lipid extract was found, as expected, to contain significant levels of numerous polar and nonpolar phospholipid species in addition to the polar ganglioside lipids described above, we evaluated the utility of the monophasic retina lipid extraction method for comprehensive lipidome analysis, compared with the conventional modified Folch retina lipid extract that is typical of the biphasic lipid extraction procedures used in most lipidomic analysis studies
Despite the presence of modest relative ion suppression effects resulting from the mixture complexity and presence of abundant phospholipids, careful appraisal of sample preparation and analysis conditions enables the effective use of monophasic lipid extraction for the evaluation of rat retina ganglioside content
Summary
Lipid extraction using a monophasic chloroform/ methanol/water mixture, coupled with functional group selective derivatization and direct infusion nano-ESI-highresolution/accurate MS, is shown to facilitate the simultaneous analysis of both highly polar and nonpolar lipids from a single retina lipid extract, including low abundance highly polar ganglioside lipids, nonpolar sphingolipids, and abundant glycerophospholipids. Quantitative comparison showed that the monophasic lipid extraction method yielded similar lipid distributions to those obtained from established “gold standard” biphasic lipid extraction methods known to enrich for either highly polar gangliosides or nonpolar lipids, respectively, with only modest relative ion suppression effects This improved lipid extraction and analysis strategy enables detailed lipidome analyses of lipid species across a broad range of polarities and abundances, from minimal amounts of biological samples and without need for multiple lipid class-specific extractions or chromatographic separation prior to analysis.—Lydic, T. Given the importance of dysregulated lipid metabolism in a number of retinal disorders, studies aimed at elucidating the role of lipids in retinal diseases would benefit from analytical strategies capable of facilitating truly comprehensive lipid analysis of both polar and nonpolar lipid classes, as well as abundant and low abundance lipids, from a single lipid extract obtained from a minimal amount of a biological sample. LC-MS has frequently been employed in lipidomic analyses, as the introduction of a chromatographic
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Free) 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
