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An improved SPE method for fractionation and identification of phospholipids
Author(s) -
Fauland Alexander,
Trötzmüller Martin,
Eberl Anita,
AfiuniZadeh Somaieh,
Köfeler Harald,
Guo Xinghua,
Lankmayr Ernst
Publication year - 2013
Publication title -
journal of separation science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.72
H-Index - 102
eISSN - 1615-9314
pISSN - 1615-9306
DOI - 10.1002/jssc.201200708
Subject(s) - chromatography , chemistry , phosphatidylethanolamine , phosphatidic acid , phospholipid , sphingomyelin , phosphatidylcholine , high performance liquid chromatography , mass spectrometry , lipidomics , formic acid , solid phase extraction , phosphatidylglycerol , sample preparation , membrane , biochemistry
This work reports an efficient and universal SPE method developed for separation and identification of phospholipids derived from complex biological samples. For the separation step, sequential combination of silica gel‐aminopropyl‐silica gel SPE cartridges is applied. This setup enables separation of phosphatidylcholine, lysophosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol, phosphatidic acid, phosphatidylinositol, phosphatidylserine, cardiolipin, and sphingomyelin into four fractions according to the polarity of their headgroups. Sample acquisition of the SPE fractions is performed by a high‐resolution LC ‐ MS system consisting of a hybrid linear IT F ourier transform ion cyclotron resonance mass spectrometer coupled to RP ‐ HPLC . The unequivocal advantage of our SPE sample preparation setup is avoidance of analyte peak overlapping in the determination step done by RP ‐ HPLC . Overlapping phospholipid signals would otherwise exert adverse ion suppression effects. An additional benefit of this method is the elimination of polar and nonpolar (e.g. neutral lipids) contaminants from the phospholipid fractions, which highly reduces contamination of the LC ‐ MS system. The method was validated with fermentation samples of organic waste, where 78 distinct phospholipid and sphingomyelin species belonging to six lipid classes were successfully identified.