Abstract
Profiling of leaf extracts from mutants of Arabidopsis with defects in lipid desaturation demonstrates the utility of collision-induced dissociation time-of-flight mass spectrometry (CID-TOF MS) for screening biological samples for fatty acid compositional alterations. CID-TOF MS uses the collision cell of a quadrupole time-of-flight mass spectrometer to simultaneously fragment all of the ions produced by an ionization source. Electrospray ionization CID-TOF MS in the negative mode can be used to analyze fatty acyl anions derived from complex lipids as well as free fatty acids. Although acyl anion yield is shown to be a function of the lipid class and the position on the glycerol backbone, acyl compositional profiles can be determined, and the TOF detector provides resolution of nominally isobaric acyl species in the profiles. Good precision is obtained when data are acquired for approximately 1 min per sample.
Highlights
Profiling of leaf extracts from mutants of Arabidopsis with defects in lipid desaturation demonstrates the utility of collision-induced dissociation time-of-flight mass spectrometry (CID-TOF collision-induced dissociation time-offlight mass spectrometry (MS)) for screening biological samples for fatty acid compositional alterations
Fatty acyl composition is most commonly analyzed by gas chromatography after derivatization to fatty acid methyl esters
The TOF analyzer provided accurate mass characterization of all intact acyl fragments in a single spectrum. Using this method as a “discovery scan,” we identified fatty acyl groups, including several oxygenated species that increased in response to wounding of the plant Arabidopsis thaliana [8]
Summary
1-15:0,2-15:0 phosphatidylethanolamine (di15:0 PE), POPC, 1-oleoyl (18:1),2-palmitoyl (16:0) phosphatidylcholine (OPPC), and 1-palmitoyl (16:0),2-oleoyl (18:1) phosphatidylethanolamine (POPE) were purchased from Avanti Polar Lipids, Inc. (Alabaster, AL). 17:0 fatty acid was purchased from NuChek Prep, Inc. (Elysian, MN). 1-18:1,2-16:1 monogalactosyldiacylglycerol (MGDG) was prepared semisynthetically [9].
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