J. Lipid Res.
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Originally published In Press as doi:10.1194/jlr.D600012-JLR200 on May 8, 2006

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Journal of Lipid Research, Vol. 47, 1865-1873, August 2006
Copyright © 2006 by American Society for Biochemistry and Molecular Biology

Methods

Characterization and direct quantitation of sphingoid base-1-phosphates from lipid extracts: a shotgun lipidomics approach

Xuntian Jiang and Xianlin Han1

Division of Bioorganic Chemistry and Molecular Pharmacology, Department of Medicine, Washington University School of Medicine, St. Louis, MO 63110

Published, JLR Papers in Press, May 8, 2006.

1 To whom correspondence should be addressed. e-mail: xianlin{at}wustl.edu

Here, we have extended shotgun lipidomics for the characterization and quantitation of sphingosine-1-phosphate (S1P) and dihydrosphingosine-1-phosphate (DHS1P) in crude lipid extracts in the presence of ammonium hydroxide by using precursor ion scanning of m/z 79.0 (corresponding to [PO3]–) in the negative-ion mode. It is demonstrated that a broad linear dynamic range for the quantitation of both S1P and DHS1P and a detection limit at low amol/µl concentration are achieved using this approach. The developed method for the quantitation of sphingoid base-1-phosphates is generally simpler and more efficient than other previously published methods. Multiple factors influencing the quantitation of sphingoid base-1-phosphates, including ion suppression, extraction efficiency, and potential overlapping with other molecular species, were examined extensively and/or are discussed. Mass levels of S1P and DHS1P in multiple biological samples, including human plasma, mouse plasma, and mouse brain tissues (e.g., cortex, cerebellum, spinal cord, and brain stem), were determined by the developed methodology. Accordingly, this technique, as a new addition to shotgun lipidomics technology, will be extremely useful for understanding the pathways of sphingolipid metabolism and for exploring the important roles of sphingoid base-1-phosphates in a wide range of physiological and pathological studies.

Supplementary key words dihydrosphingosine-1-phosphate • electrospray ionization mass spectrometry • brain tissues • sphingolipid metabolism


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Copyright © 2006 by the American Society for Biochemistry and Molecular Biology.