title: Rapid nanoscale quantitative analysis of plant sphingolipid long-chain bases by GC-MS.
abstract: In eukaryotic organisms, sphingolipids are major structural lipids of biological membranes and perform additional essential functions as signalling molecules. While long-chain bases (LCB), the common precursor to all sphingolipid classes, is represented by only one major molecular species in animals and fungi, up to nine LCB have been found in plants. In the absence of genuine plant sphingolipid references required for proper quantification, we have reinvestigated and optimized a protocol destined to the quantification of total plant LCB that relies on the use of gas chromatography-mass spectrometry (GC-MS). This rapid three-step protocol sequentially involves (1) the release of LCB from biological samples using barium hydroxide solution, (2) their oxidation into aldehydes by metaperiodate, and (3) the subsequent identification/quantification of these aldehydes by GC-MS. It is simple and reliable and enables separation of aldehydes upon their stero-specificity. It further enables the quantification of total LCB from a wide variety of samples including yeast and animal cell cultures.
authors.affiliation: Laboratoire de Biogenèse Membranaire, UMR 5200 CNRS-Université Victor Segalen Bordeaux 2, Bordeaux, France. cacasjl@yahoo.fr
authors.forename: Jean-Luc
authors.initials: JL
authors.lastname: Cacas
authors.forename: Su
authors.initials: S
authors.lastname: Melser
authors.forename: Frédéric
authors.initials: F
authors.lastname: Domergue
authors.forename: Jérôme
authors.initials: J
authors.lastname: Joubès
authors.forename: Brice
authors.initials: B
authors.lastname: Bourdenx
authors.forename: Jean-Marie
authors.initials: JM
authors.lastname: Schmitter
authors.forename: Sébastien
authors.initials: S
authors.lastname: Mongrand
mesh_terms: D017609:Barium Compounds / Q000737:chemistry; D008401:Gas Chromatography-Mass Spectrometry / Q000191:economics / Q000379:methods*; D010084:Oxidation-Reduction; D010936:Plant Extracts / Q000032:analysis* / Q000302:isolation & purification; D010944:Plants / Q000737:chemistry*; D012680:Sensitivity and Specificity; D013107:Sphingolipids / Q000032:analysis* / Q000302:isolation & purification; D013997:Time Factors
chemical_list: D017609:Barium Compounds; D010936:Plant Extracts; D013107:Sphingolipids; C012766:barium hydroxide