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Screening of stationary phase selectivities for global lipid profiling by ultrahigh performance supercritical fluid chromatography

Al Hamimi, Said LU ; Sandahl, Margareta LU ; Armeni, Marina; Turner, Charlotta LU and Spégel, Peter LU (2018) In Journal of Chromatography A 1548. p.76-82
Abstract

The performance of seven sub-2-μm particle packed columns (2-picolylamine, 2-PIC; charged surface hybrid fluoro-phenyl, CSH-FP; high strength silica C18 SB, HSS-C18; diethylamine, DEA; 1-aminoanthracene, 1-AA; high density diol and ethylene bridged hybrid; BEH) was examined for lipid separation in ultra-high performance supercritical fluid chromatography (UHPSFC) coupled to quadrupole time-of-flight mass spectrometry. Based on the results of the column screening a method for profiling of multiple lipid species from the major lipid classes was developed. Stationary phases containing β-hydroxy amines, i.e. 1-AA, DEA and 2-PIC, yielded strong retention and poor peak shapes of zwitterionic lipids with primary amine groups, such... (More)

The performance of seven sub-2-μm particle packed columns (2-picolylamine, 2-PIC; charged surface hybrid fluoro-phenyl, CSH-FP; high strength silica C18 SB, HSS-C18; diethylamine, DEA; 1-aminoanthracene, 1-AA; high density diol and ethylene bridged hybrid; BEH) was examined for lipid separation in ultra-high performance supercritical fluid chromatography (UHPSFC) coupled to quadrupole time-of-flight mass spectrometry. Based on the results of the column screening a method for profiling of multiple lipid species from the major lipid classes was developed. Stationary phases containing β-hydroxy amines, i.e. 1-AA, DEA and 2-PIC, yielded strong retention and poor peak shapes of zwitterionic lipids with primary amine groups, such as phosphatidylserines, phosphatidylethanolamines and its lyso forms. The BEH and HSS-C18 columns showed strong retention of polar and nonpolar lipids, respectively. The Diol column retained the majority of major lipid classes and also produced symmetric peaks. In addition, this column also produced the highest resolution within and between major lipid classes. An injection solvent composed of methanol:chloroform (1:2, v:v) and the addition of 20 mM ammonium formate in the mobile phase improved chromatographic separation and mass spectrometry detection in comparison to ammonium acetate or absence of additive. Finally, chromatographic and mass spectrometric parameters were optimized for the Diol column using a design of experiments approach. The separation mechanism on the Diol column depended on the lipid functionality and the length and degree of unsaturation of the acyl groups. The developed method could resolve 18 lipid classes and multiple lipids within each class, from blood serum and brain tissue in 11 min.

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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Blood serum, Design of experiments, Lipidomics, Mass spectrometry
in
Journal of Chromatography A
volume
1548
pages
7 pages
publisher
Elsevier
external identifiers
  • scopus:85044115810
ISSN
0021-9673
DOI
10.1016/j.chroma.2018.03.024
language
English
LU publication?
yes
id
93f83339-6b7d-46ef-8489-f5117f793345
date added to LUP
2018-04-09 13:58:48
date last changed
2019-03-19 03:52:27
@article{93f83339-6b7d-46ef-8489-f5117f793345,
  abstract     = {<p>The performance of seven sub-2-μm particle packed columns (2-picolylamine, 2-PIC; charged surface hybrid fluoro-phenyl, CSH-FP; high strength silica C18 SB, HSS-C<sub>18</sub>; diethylamine, DEA; 1-aminoanthracene, 1-AA; high density diol and ethylene bridged hybrid; BEH) was examined for lipid separation in ultra-high performance supercritical fluid chromatography (UHPSFC) coupled to quadrupole time-of-flight mass spectrometry. Based on the results of the column screening a method for profiling of multiple lipid species from the major lipid classes was developed. Stationary phases containing β-hydroxy amines, i.e. 1-AA, DEA and 2-PIC, yielded strong retention and poor peak shapes of zwitterionic lipids with primary amine groups, such as phosphatidylserines, phosphatidylethanolamines and its lyso forms. The BEH and HSS-C<sub>18</sub> columns showed strong retention of polar and nonpolar lipids, respectively. The Diol column retained the majority of major lipid classes and also produced symmetric peaks. In addition, this column also produced the highest resolution within and between major lipid classes. An injection solvent composed of methanol:chloroform (1:2, v:v) and the addition of 20 mM ammonium formate in the mobile phase improved chromatographic separation and mass spectrometry detection in comparison to ammonium acetate or absence of additive. Finally, chromatographic and mass spectrometric parameters were optimized for the Diol column using a design of experiments approach. The separation mechanism on the Diol column depended on the lipid functionality and the length and degree of unsaturation of the acyl groups. The developed method could resolve 18 lipid classes and multiple lipids within each class, from blood serum and brain tissue in 11 min.</p>},
  author       = {Al Hamimi, Said and Sandahl, Margareta and Armeni, Marina and Turner, Charlotta and Spégel, Peter},
  issn         = {0021-9673},
  keyword      = {Blood serum,Design of experiments,Lipidomics,Mass spectrometry},
  language     = {eng},
  month        = {05},
  pages        = {76--82},
  publisher    = {Elsevier},
  series       = {Journal of Chromatography A},
  title        = {Screening of stationary phase selectivities for global lipid profiling by ultrahigh performance supercritical fluid chromatography},
  url          = {http://dx.doi.org/10.1016/j.chroma.2018.03.024},
  volume       = {1548},
  year         = {2018},
}