Journal
LIPIDS
Volume 48, Issue 3, Pages 307-318Publisher
SPRINGER HEIDELBERG
DOI: 10.1007/s11745-013-3760-z
Keywords
Lipidomics; Human-brain; Ceramide; Sphingomyelin; Phosphatidylcholine; Phosphatidylethanolamine; Phosphatidylserine; Sterol
Funding
- National Health and Medical Research Council of Australia
- University of New South Wales
- Neuroscience Research Australia
- Australian National Health and Medical Research Council (NHMRC) [APP1008307]
- Australian Research Council [FT110100249, FT0991986]
- NHMRC [630434]
- Australian Research Council [FT0991986] Funding Source: Australian Research Council
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We have developed a protocol suitable for high-throughput lipidomic analysis of human brain samples. The traditional Folch extraction (using chloroform and glass-glass homogenization) was compared to a high-throughput method combining methyl-tert-butyl ether (MTBE) extraction with mechanical homogenization utilizing ceramic beads. This high-throughput method significantly reduced sample handling time and increased efficiency compared to glass-glass homogenizing. Furthermore, replacing chloroform with MTBE is safer (less carcinogenic/toxic), with lipids dissolving in the upper phase, allowing for easier pipetting and the potential for automation (i.e., robotics). Both methods were applied to the analysis of human occipital cortex. Lipid species (including ceramides, sphingomyelins, choline glycerophospholipids, ethanolamine glycerophospholipids and phosphatidylserines) were analyzed via electrospray ionization mass spectrometry and sterol species were analyzed using gas chromatography mass spectrometry. No differences in lipid species composition were evident when the lipid extraction protocols were compared, indicating that MTBE extraction with mechanical bead homogenization provides an improved method for the lipidomic profiling of human brain tissue.
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