title: High-throughput phospholipid quantitation in mammalian cells using matrix-assisted laser desorption ionization-time of flight mass spectrometry with N-trifluoroacetyl-phosphatidylethanolamine as internal standard.
abstract: A high-throughput method is described for quantitative analysis of phospholipids. The method comprises extraction of lipids, addition of the internal standard N-trifluoroacetyl-phosphatidylethanolamine, and final analysis using matrix-assisted laser desorption ionization mass spectrometry. Quantitative data are obtained by calibration directly in the sample matrix. Calibration with one phosphatidylcholine was found sufficient for quantification of all major phosphatidylcholines tested. The method is very sensitive, has broad application, and is easily applicable to any biological sample. The detection limit for phosphatidylcholines was clearly below 2 μg on the spot, requiring less than 4000 cells corresponding to about 1.6 μg cell dry mass.
authors.affiliation: Biochemical Engineering Institute, Saarland University, D-66123 Saarbrücken, Germany.
authors.forename: Jens
authors.initials: J
authors.lastname: Niklas
authors.forename: Klaus
authors.initials: K
authors.lastname: Hollemeyer
authors.forename: Elmar
authors.initials: E
authors.lastname: Heinzle
mesh_terms: D000818:Animals; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D057166:High-Throughput Screening Assays / Q000379:methods*; D010714:Phosphatidylethanolamines / Q000032:analysis*; D012015:Reference Standards; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D010714:Phosphatidylethanolamines; C068825:N-trifluoroacetyl-1,2-dioleoylphosphatidylethanolamine; C483858:phosphatidylethanolamine