High-throughput lipid extraction for the analysis of human brain lipids - Bertin Instruments

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High-throughput lipid extraction for the analysis of human brain lipids

Sources: Illawarra Health and Medical Research Institute, University of Wollongong, Australia.

Context

Traditional lipid extraction techniques are the bottleneck for modern shotgun lipidomic studies. To overcome this problem, protocol comparisons were made between the traditional Folch extraction (using chloroform and glass-glass homogenization) and a high-throughput method combining methyl-tert-butyl ether (MTBE) with mechanical homogenization (Precellys-24, Bertin Technologies) [1].

 

[1] Sarah K. Abbott et al. An Improved High-Throughput Lipid Extraction Method for the Analysis of Human Brain Lipids, Lipids ISSN 0024-4201 Volume 48 Number 3 Lipids (2013) 48:307-318 DOI 10.1007/s11745-013-3760-z

Results

Traditional lipid extraction techniques are the bottleneck for  modern  shotgun lipidomic studies.  To  overcome this problem, protocol comparisons were made between the  traditional  Folch extraction  (using  chloroform  and glass-glass  homogenization)  and  a  high-throughput method  combining  methyl-tert-butyl  ether  (MTBE)  with mechanical  homogenization  (Precellys-24,  Bertin Technologies) [1].

 

03712-810-DU076

Figure 1: Representative spectra of human  occipital  cortex comparing  Bead  – MTBE  to  Glass  – Chloroform  (PC  head group scan: precursor ion scan m/z 184.1)

This  high-throughput  Bead-MTBE  protocol  improves upon  traditional  lipid  extraction  methods  as  it  is  safer (less  carcinogenic/toxic) and  much  more efficient.  The Bead-MTBE  protocol  is  approximately  four  times quicker than Glass-Chloroform in the homogenization of 24  samples  (i.e.1  vs.  4  h),  with  the  additional  benefit being that tissue aliquots can be weighted directly into the  Precellys tubes  prior  to  homogenization  (thus reducing  double-handling  times).  The  lower  density  of MTBE  further  enhances  the  lipid  extraction  procedure (by  dissolving  lipids  in  the  upper  phase)  and  is  also better  for  the  potential  incorporation  of  robotics  to further streamline lipidomic studies.

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