Shotgun lipidomics of phosphoethanolamine-containing lipids in biological samples after one-step in situ derivatization

Autor: Xianlin Han, Kui Yang, Hua Cheng, Kora N. Fikes, Richard W. Gross
Jazyk: angličtina
Rok vydání: 2005
Předmět:
Zdroj: Journal of Lipid Research, Vol 46, Iss 7, Pp 1548-1560 (2005)
Druh dokumentu: article
ISSN: 0022-2275
DOI: 10.1194/jlr.D500007-JLR200
Popis: 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 infused directly into the ion source of an electrospray ionization 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 attomoles (amoles) per microliter for PE and lysoPE analysis was readily achieved using this technique (at least a 100-fold improvement from our previous method) with a >15,000-fold dynamic range. Through intrasource separation and multidimensional mass spectrometry 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 farther into the low-abundance regime of cellular lipidomes.
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