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

被引：78

作者：

Abbott, Sarah K. ^{[1
,2
]}

Jenner, Andrew M. ^{[1
,2
]}

Mitchell, Todd W. ^{[1
,3
]}

Brown, Simon H. J. ^{[1
,3
]}

Halliday, Glenda M. ^{[4
,5
]}

Garner, Brett ^{[1
,2
]}

机构：

[1] Univ Wollongong, Illawarra Hlth & Med Res Inst, Wollongong, NSW 2522, Australia

[2] Univ Wollongong, Sch Biol Sci, Wollongong, NSW 2522, Australia

[3] Univ Wollongong, Sch Hlth Sci, Wollongong, NSW 2522, Australia

[4] Neurosci Res Australia, Sydney, NSW 2031, Australia

[5] Univ New S Wales, Sydney, NSW 2031, Australia

来源：

LIPIDS | 2013年 / 48卷 / 03期

基金：

澳大利亚国家健康与医学研究理事会; 澳大利亚研究理事会; 英国医学研究理事会;

关键词：

Lipidomics; Human-brain; Ceramide; Sphingomyelin; Phosphatidylcholine; Phosphatidylethanolamine; Phosphatidylserine; Sterol; ALZHEIMERS-DISEASE; FATTY-ACIDS; SHOTGUN LIPIDOMICS; MASS-SPECTROMETRY; DEFICIENCY; TISSUE; QUANTIFICATION; IDENTIFICATION; SPHINGOMYELIN; PHOSPHOLIPIDS;

D O I：

10.1007/s11745-013-3760-z

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

070307 [化学生物学]; 071010 [生物化学与分子生物学];

摘要：

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.

引用

页码：307 / 318

页数：12

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