Alkaline methanolysis of lipid extracts extends shotgun lipidomics analyses to the low-abundance regime of cellular sphingolipids

Xuntian Jiang, Hua Cheng, Kui Yang, Richard W. Gross, Xianlin Han

Research output: Contribution to journalArticlepeer-review

60 Scopus citations

Abstract

Sphingolipids that contain a sphingoid base are composed of hundreds to thousands of distinct compounds, many of which serve as lipid regulators of biological functions. The global analysis of the large number of low-abundance sphingolipid molecular species has been hampered in many cases by the sphingolipid molecular species being overwhelmed by the quantity of other classes of lipid (e.g., glycerophospholipid) molecular species present, thereby imposing severe restrictions on the dynamic range of their measurement using shotgun lipidomics. Herein, we developed a facile approach in which the sphingolipids of cellular extracts were dramatically enriched by direct alkaline methanolysis of lipid extracts followed by extraction to remove the large majority of other endogenous lipid classes. Through direct infusion of the resultant enriched solution, we identified and quantitated a variety of very-low-abundance sphingolipid classes (e.g., sphingosine, psychosine, and lysosphingomyelin) and molecular species (e.g., sphingomyelin) using electrospray ionization mass spectrometry (i.e., shotgun sphingolipidomics). Accordingly, through utilization of these facile enrichment techniques, direct penetrance into the sphingolipidomes has been greatly extended, facilitating new insights into their metabolism and signaling functions in biological systems.

Original languageEnglish (US)
Pages (from-to)135-145
Number of pages11
JournalAnalytical Biochemistry
Volume371
Issue number2
DOIs
StatePublished - Dec 15 2007
Externally publishedYes

Keywords

  • Electrospray ionization mass spectrometry
  • Lysosphingomyelin
  • Psychosine
  • Shotgun lipidomics
  • Shotgun sphingolipidomics
  • Sphingolipidome
  • Sphingomyelin
  • Sphingosine

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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