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

Vitali Matyash, Gerhard Liebisch, Teymuras V. Kurzchalia, Andrej Shevchenko, and Dominik Schwudke

Max Planck Institute of Molecular Cell Biology and Genetics, Dresden 01307

Corresponding Author: schwudke{at}mpi-cbg.de

Accurate profiling of lipidomes relies upon 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-buthylether (MTBE) extraction allows faster and cleaner lipid recovery and is well suited for automated shotgun profiling. Because of MTBE low density, lipid-containing organic phase forms the upper layer during phase separation, which simplifies its collection and minimizes dripping losses. Non-extractable matrix forms a dense pellet at the bottom of the extraction tube and easily removed by centrifuging. Rigorous testing demonstrated that MTBE protocol delivers similar or better recoveries of species of most all major lipid classes, compared to “golden standard” Folch and Bligh and Dyer recipes.

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