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A more recent version of this article appeared on November 1, 2003

Papers In Press, published online ahead of print August 16, 2003
J. Lipid Res., doi:10.1194/jlr.D300020-JLR200
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Submitted on June 27, 2003
Revised on August 16, 2003
Accepted on August 11, 2003

Charting molecular composition of phosphatidylcholines by fatty acid scanning and ion trap MS3 fragmentation

Kim Ekroos, Christer S. Ejsing, Ute Bahr, Michael Karas, Kai Simons, and Andrej Shevchenko

MPI of Molecular Cell Biology and Genetics, Dresden 01307

Corresponding Author: shevchenko{at}mpi-cbg.de

The molecular composition of phosphatidylcholines (PCs) in total lipid extracts was characterized by a combination of multiple precursor ion scanning on a hybrid quadrupole time-of-flight mass spectrometer and MS3 fragmentation on an ion trap mass spectrometer. Precursor ion spectra for 50 acyl anion fragments of fatty acids (Fatty Acid Scanning, or FAS) acquired in parallel increased the specificity and the dynamic range of the detection of PCs and identified the fatty acid moieties in individual PC species. Subsequent analysis of detected PC peaks by MS3 fragmentation on an ion trap mass spectrometer quantified the relative amount of their positional isomers, thus providing the most detailed and comprehensive characterization of the molecular composition of the pool of PCs at the low picomole level. The method is vastly simplified compared to conventional approaches and does not require preliminary separation of lipid classes or individual molecular species, enzymatic digestion or chemical derivatization. The approach was validated by the comparative analysis of the molecular composition of PCs from human red blood cells. In the total lipid extract of MDCK II cells, we detected 46 PC species with unique fatty acid composition and demonstrated that the presence of positional isomers almost doubled the total number of individual molecular species.


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