J. Lipid Res.
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH
QUICK SEARCH:   [advanced]


     

A more recent version of this article appeared on November 1, 2003

Papers In Press, published online ahead of print August 1, 2003
J. Lipid Res., doi:10.1194/jlr.D300025-JLR200
This Article
Right arrow Full Text (Accepted Manuscript)
Right arrow All Versions of this Article:
D300025-JLR200v1
44/11/2209    most recent
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Lieser, B.
Right arrow Articles by Schmitz, G.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Lieser, B.
Right arrow Articles by Schmitz, G.
Social Bookmarking
Add to CiteULike   Add to Complore   Add to Connotea   Add to Del.icio.us   Add to Digg   Add to Reddit   Add to Technorati  
What's this?

Submitted on July 9, 2003
Revised on August 1, 2003
Accepted on July 25, 2003

Quantification of sphingosine and sphinganine from crude lipid extracts by HPLC-electrospray ionization tandem mass spectrometry

Bernd Lieser, Gerhard Liebisch, Wolfgang Drobnik, and Gerd Schmitz

Institute for Clinical Chemistry and Laboratory Medicine, University of Regensburg, Regensburg 93042

Corresponding Author: gerd.schmitz{at}klinik.uni-regensburg.de

Sphingosine (SPH) comprises the backbone of sphingolipids and is known as a second messenger involved in the modulation of cell growth, differentiation and apoptosis. The currently available methods for the quantification of SPH are in part complicated, time consuming, insensitive or unselective. Therefore a fast and convenient methodology for the quantification of SPH and the biosynthetic intermediate sphinganine (SPA) was developed. The method is based on an HPLC separation coupled to electrospray ionization tandem mass spectrometry (ESI-MS/MS). Quantitation is achieved by use of a constant concentration of a non-naturally occurring internal standard C17-SPH, together with a calibration curve established by spiking different concentrations of naturally occurring sphingoid bases. SPH and SPA co-eluted with C17-SPH, which allows an accurate correction of the analyte response. Interference of the SPH+2 isotope with SPA quantification was corrected by an experimentally determined factor. The limits of detection were 9 fmol for SPH and 21 fmol for SPA. The overall coefficients of variation (CV) were 8 % and 13 % for SPH and SPA, respectively. The developed LC-MS/MS methodology with an analysis time of 3.5 min, simple sample preparation and automated data analysis allows high throughput quantification of sphingoid bases from crude lipid extracts and is a valuable tool for studies of cellular sphingolipid metabolism and signaling.


Add to CiteULike CiteULike   Add to Complore Complore   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us   Add to Digg Digg   Add to Reddit Reddit   Add to Technorati Technorati    What's this?


This article has been cited by other articles:


Home page
 Mol. Biol. CellHome page
S. C.D. van IJzendoorn, J. M. van der Wouden, G. Liebisch, G. Schmitz, and D. Hoekstra
Polarized Membrane Traffic and Cell Polarity Development Is Dependent on Dihydroceramide Synthase-Regulated Sphinganine Turnover
Mol. Biol. Cell, September 1, 2004; 15(9): 4115 - 4124.
[Abstract] [Full Text] [PDF]


HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH
 All ASBMB Journals   Journal of Biological Chemistry 
 Molecular and Cellular Proteomics   ASBMB Today 
Copyright © 2003 by the American Society for Biochemistry and Molecular Biology.