J. Lipid Res. Neurobiology of Lipids (ISSN1683-5506)
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A more recent version of this article appeared on July 1, 2005

Papers In Press, published online ahead of print April 16, 2005
J. Lipid Res., doi:10.1194/jlr.D500007-JLR200
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Submitted on February 28, 2005
Revised on March 31, 2005
Accepted on April 3, 2005

Shotgun lipidomics of phosphoethanolamine-containing lipid molecular species in biological samples after one-step in situ derivatization

Xianlin Han, Kui Yang, Hua Cheng, Kora Fikes, and Richard W. Gross

Dept of Internal Medicine, Washington University School of Medicine, St. Louis, MO 63110

Corresponding Author: xianlin{at}wustl.edu

This article presents a novel methodology for the analysis of ethanolamine glycerophospholipid (PE) and lysoPE molecular species directly from lipid extracts of biological samples. Through brief treatment of lipid extracts with fluorenylmethoxylcarbonyl (Fmoc) chloride, PE and lysoPE species were selectively derivatized to their corresponding carbamates. The reaction solution was directly infused into the ion source of an ESI mass spectrometer after appropriate dilution. The facile loss of the Fmoc moiety dramatically enhanced the analytic sensitivity and allowed the identification and quantitation of low-abundance molecular species. A detection limitation of amol/µl for PE and lysoPE analysis was readily achieved using this technique (at least a 100-fold improvement from our previous method) with over a 15,000-fold dynamic range. Through intrasource separation and multi-dimensional MS array analysis of derivatized species, marked improvements in signal-to-noise ratio, molecular species identification, and quantitation can be realized. The procedure is both simple and effective and can be extended to analyze many other lipid classes or other cellular metabolites by adjustments in specific derivatization conditions. Thus through judicious derivatization, a new dimension exploiting specific functional reactivities in each lipid class can be used in conjunction with shotgun lipidomics to penetrate further into the low-abundance regime of cellular lipidomes.


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