Determination of content and fatty acid composition of unlabeled phosphoinositide species by thin-layer chromatography and gas chromatography.
Recent advances in research on the physiological roles of phosphoinositides in eukaryotic organisms indicate a need to distinguish molecular phosphoinositide species on the basis of their characteristic head groups as well as their glycerolipid moieties. Accurate identification of phosphoinositide species in biological samples poses an analytical challenge, because structurally similar inositol phosphate head groups must be resolved, as must lipid-associated fatty acids. Although intact phosphoinositide species have been successfully analyzed, such analyses employ state-of-the-art liquid chromatography/mass spectrometry and require expensive equipment not accessible to many researchers. Described here is a cost-efficient and reliable alternative developed by adaptation of a combination of classic methods for lipid analysis, thin-layer chromatography and gas chromatography.
König
Sabine
S
Department of Plant Biochemistry, Albrecht-von-Haller-Institute for Plant Sciences, Georg-August-University Göttingen, 37077 Göttingen, Germany.
Hoffmann
Mareike
M
Mosblech
Alina
A
Heilmann
Ingo
I
D017360:Arabidopsis / Q000737:chemistry; D002849:Chromatography, Gas / Q000379:methods*; D002855:Chromatography, Thin Layer / Q000379:methods*; D005227:Fatty Acids / Q000032:analysis*; D007536:Isomerism; D010716:Phosphatidylinositols / Q000032:analysis* / Q000737:chemistry / Q000302:isolation & purification; D018515:Plant Leaves / Q000737:chemistry; D012441:Saccharomyces cerevisiae / Q000737:chemistry; D014026:Nicotiana / Q000737:chemistry
D005227:Fatty Acids; D010716:Phosphatidylinositols