, Volume 48, Issue 3, pp 307–318 | Cite as

An Improved High-Throughput Lipid Extraction Method for the Analysis of Human Brain Lipids

  • Sarah K. AbbottEmail author
  • Andrew M. Jenner
  • Todd W. Mitchell
  • Simon H. J. Brown
  • Glenda M. Halliday
  • Brett Garner


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.


Lipidomics Human-brain Ceramide Sphingomyelin Phosphatidylcholine Phosphatidylethanolamine Phosphatidylserine Sterol 



Butylated hydroxytoluene




Collision energy






Choline glycerophospholipid


Coefficient of variation


Collision cell exit potential


Declustering potential


Entrance potential


Electrospray ionization mass spectrometry


Ethanolamine glycerophospholipid


Gas chromatography mass spectrometry


Methyl-tert-butyl ether


Multiple reaction monitoring


Neutral loss


Precursor ion









Adult brain tissue was received from the Sydney Brain Bank at Neuroscience Research Australia which is supported by the National Health and Medical Research Council of Australia, the University of New South Wales and Neuroscience Research Australia. This research was supported by a project grant (APP1008307) from the Australian National Health and Medical Research Council (NHMRC) awarded to GMH and BG. TWM and BG are supported by Australian Research Council Future Fellowships (FT110100249 and FT0991986, respectively). GMH is supported by an NHMRC Principal Research Fellowship (630434).

Conflict of interest

All authors disclose that there are no conflicts of interest.


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Copyright information

© AOCS 2013

Authors and Affiliations

  • Sarah K. Abbott
    • 1
    • 2
    Email author
  • Andrew M. Jenner
    • 1
    • 2
  • Todd W. Mitchell
    • 1
    • 3
  • Simon H. J. Brown
    • 1
    • 3
  • Glenda M. Halliday
    • 4
  • Brett Garner
    • 1
    • 2
  1. 1.Illawarra Health and Medical Research InstituteUniversity of WollongongWollongongAustralia
  2. 2.School of Biological SciencesUniversity of WollongongWollongongAustralia
  3. 3.School of Health SciencesUniversity of WollongongWollongongAustralia
  4. 4.Neuroscience Research Australia and the University of New South WalesSydneyAustralia

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