One Two
Sie sind hier: Startseite Molekulare Biotechnologie Lipidomics Platform

Lipidomics Platform

The IMBIO Institute, AG Dörmann, provides a lipid analytical platform for the campus of the University of Bonn in the framework of the SFB645. We measure a large variety of lipids in a quantitative manner employing state-of-the art analytical equipment, including fluorescence HPLC, gas chromatography (GC), GC-MS and Q-TOF (quadrupole time of flight) mass spectrometry (Figure 1).


hplcgcmsqtof

Figure 1: Equipment of the lipidomics platform at the IMBIO Institute, University of Bonn
From left to right: Agilent 1100 HPLC, Agilent GC and GC-MS, Agilent Q-TOF (quadrupole time-of flight) mass spectrometer equipped with Agilent Chipcube nanospray, APCI and ESI source.

If you are interested in lipid measurements at the lipidomics facility of University of Bonn, please check the class(es) of lipids you are interested in (this page). To cover the expenses for solvents, LC columns, and parts for chromatography and mass spectrometry, we ask for a contribution according to the table shown on this page (costs). Protocols for lipid extraction are available (boiling-water-protocol) or (acidic chloroform-methanol extraction).

For questions please contact:

Head of Department and Managing Director of IMBIO

Prof. Dr. Peter Dörmann    doermann@uni-bonn.de

 

Lipidomics Service Team

Dr. Katharina Gutbrod    k.gutbrod@uni-bonn.de

Helga Peisker                  h.peisker@uni-bonn.de

 

Lipids measured by the lipidomics platform (Letters in brackets indicate cost calculation)

Lipid isolation (A)
Glycerolipids are isolated from leaves, bacteria and fungi using the boiling-water protocol, or from animal tissues or from plant roots with acidic chloroform-methanol extraction.

Solid phase extraction (SPE) on silica columns (B)
SPE on silica columns is used to separate neutral lipids (triacylglycerol, diacylglycerol), glycolipids (galactolipids, hexosylceramides), phospholipids and lyso-phospholipids, and for the separation of sterol lipid classes from plants.

Phospholipids/Galactolipids (membrane glycerolipids) from plants, bacteria and fungi (A, C)
Phospholipid and galactolipid  from plants, bacteria and fungi are isolated using the boiling-water protocol or the acidic chloroform-methanol extraction and measured by tandem mass spectrometry (Q-TOF MS/MS). Lipids are quantified by MS/MS experiments with internal standards following the strategy developed by Ruth Welti (Kansas State University) (also see: Gasulla et al., 2013).

Lyso-phospholipids (A, B, C)
After lipid isolation, lyso-phospholipids (lyso-PC) are purified by SPE on silica columns prior to measurement by direct infusion Q-TOF mass spectrometry.

Sphingolipids from animals (please contact us)
The sphingolipid composition from animals (ceramide, hexosylceramide, sphingomyelin, gangliosides,sulfatides) can be measured after alkaline hydrolysis of phospholipids. Neutral and acidic sphingolipids are separated by solid phase extraction and quantified by direct infusion Q-TOF MS/MS.

Sphingolipids from Plants (please contact us)
Plant sphingolipids in crude lipid extracts are  purified by alkaline hydrolysis of phospholipids and galactolipids.  Glucosylceramides from plants can be measured by direct infusion with the Q-TOF mass spectrometer after SPE on silica columns.The full spectrum of sphingolipids (ceramide, glucosylceramide, sphingobase, GIPC) can be measured by LC-MS/MS following a protocol developed by Jonathan Markham (University of Lincoln Nebraska).

Triacylglycerol, Diacylglycerol (A, B, C)
Triacylglycerol and diacylglycerol are isolated from the sample using the boiling-water protocol or the acidic chloroform-methanol extraction and then separated from polar lipids by SPE on silica columns. Triacylglycerol and diacylglycerol are quantified by direct infusion mass spectrometry (Q-TOF) (Lippold et al., 2012; vom Dorp et al., 2013).

Fatty acids (E, F)
Total fatty acids in biological samples can be quantified after conversion of all acyl groups into fatty acid methyl esters. The methyl esters are quantified by gas chromatography (GC) with flame ionization detector using pendadecanoic acid (15:0) as internal standard. For structural identification, fatty acid methyl esters are analyzed by gas chromatography-mass spectrometry (GC-MS).

Sterol lipids (A, B, C, E, F)
Free sterols can be measured by GC-MS after silylation, and total sterols (free and conjugated) sterols can be measured in the same way after acidic/alkaline hydrolysis and silylation (A, E, F)
In order to analyze the full spectrum of free and conjugated sterols, the sterol lipid classes are separated by SPE on silica columns  into a polar fraction (only present in plants and some fungi: sterol glucosides, acylated sterol glucosides) and a non-polar fraction (free sterols, sterol esters). These fractions are measured separately by direct infusion Q-TOF MS (A, B, C). Free sterols need to be derivatized e.g. with chlorobetainyl chloride prior to quantification (Wewer et al., 2010).


References

Artikelaktionen