Lipid extraction using MTBE

From LipidomicsWiki

Description

• liquid-liquid extraction using MTBE (methyl-tert-butyl ether)

Protocol

• aqueous samples are used for lipid extraction including cell homogenates, homogenized tissue, plasma
• to 0.2 volume parts aqueous sample 1.5 parts of methanol are added and mixed
• 5 ml of MTBE was added
• the mixture was incubated for 1h at room temperature in a shaker
• add 1.25 volume part water for phases separation
• after mixing the sample is centrifuged at 1.000g for 10 min
• the upper MTBE phase containing the lipids is collected
• the lower is re-extracted with 2 volume parts of MTBE/methanol/water (10/3/2.5, v/v/v)
• the upper MTBE phase collected
• the combined MTBE phases were dried

Reference

• Matyash V, Liebisch G, Kurzchalia TV, Shevchenko A, Schwudke D. J Lipid Res. 2008 May;49(5):1137-46.