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Originally published In Press as doi:10.1194/jlr.D700041-JLR200 on February 16, 2008

Papers In Press, published online ahead of print May 1, 2008
J. Lipid Res., doi:10.1194/jlr.D700041-JLR200
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Journal of Lipid Research, Vol. 49, 1137-1146, May 2008
Copyright © 2008 by American Society for Biochemistry and Molecular Biology

Methods

Lipid extraction by methyl-tert-butyl ether for high-throughput lipidomics*,boxs

Vitali Matyash*, Gerhard Liebisch{dagger}, Teymuras V. Kurzchalia*, Andrej Shevchenko1,* and Dominik Schwudke1,*

* Max Planck Institute of Molecular Cell Biology and Genetics, 01307 Dresden, Germany
{dagger} Institute of Clinical Chemistry and Laboratory Medicine, University of Regensburg, 93053 Regensburg, Germany

* This work was funded, in part, by Grant SFB TR13 from the Deutsche Forschungsgemeinschaft to T.V.K. and A.S. (projects B2 and D1, respectively) and by a Human Frontier Science Program grant to T.V.K.

boxs The online version of this article (available at http://www.jlr.org) contains supplementary data in the form of four figures.

Published, JLR Papers in Press, February 16, 2008.

1 To whom correspondence should be addressed. e-mail: shevchenko{at}mpi-cbg.de (A.S.); schwudke{at}mpi-cbg.de (D.S.)

Accurate profiling of lipidomes relies upon the quantitative and unbiased recovery of lipid species from analyzed cells, fluids, or tissues and is usually achieved by two-phase extraction with chloroform. We demonstrated that methyl-tert-butyl ether (MTBE) extraction allows faster and cleaner lipid recovery and is well suited for automated shotgun profiling. Because of MTBE's low density, lipid-containing organic phase forms the upper layer during phase separation, which simplifies its collection and minimizes dripping losses. Nonextractable matrix forms a dense pellet at the bottom of the extraction tube and is easily removed by centrifugation. Rigorous testing demonstrated that the MTBE protocol delivers similar or better recoveries of species of most all major lipid classes compared with the "gold-standard" Folch or Bligh and Dyer recipes.

Supplementary key words mass spectrometry • sample preparation • direct infusion • nano-ESI-MS


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