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A more recent version of this article appeared on August 1, 2006

Papers In Press, published online ahead of print May 8, 2006
J. Lipid Res., doi:10.1194/jlr.D600012-JLR200
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Submitted on March 31, 2006
Revised on May 3, 2006
Accepted on May 8, 2006

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

Xuntian Jiang and Xianlin Han

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

Corresponding Author: xianlin{at}wustl.edu

Herein, 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 DH S1P and a detection limit at low amol/µl concentration are achieved by using this approach. The developed method for quantitation of sphingoid base-1-phosphates is generally simpler and more efficient than other previously published methods. Multiple factors influencing quantitation of sphingoid base-1-phosphates, including ion suppression, extraction efficiency, and potential overlapping with other molecular species, were extensively examined and/or 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 the 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.


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