The Lipidomics/Metabolomics Facility offers the analysis of polar metabolites and lipids in biological samples using state-of-the-art mass spectrometric techniques and data processing software. The service also includes help with the experimental design and data interpretation.
Our services focus on the following areas:
1. Liquid Chromatography / Tandem Mass Spectrometry (LC/MS/MS)
The LC/MS/MS approach includes the LC pre-separation of the metabolite/lipid extract of the biological sample prior to the MS detection. We use LC/MS/MS for targeted as well as for non-targeted analyses.
In the targeted approach, only a pre-defined set of metabolites is selectively detected and analyzed. LC-MS methods are specifically developed and optimized for the metabolites of interest. Targeted metabolomics enables the accurate quantification of the selected metabolites in the sample and the detection of even slight differences in metabolite amounts between two samples.
The CECAD Lipidomics/Metabolomics Facility offers the targeted quantification of the following lipid/metabolite classes by LC/MS/MS:
Non-targeted metabolomics is an unbiased approach to get a global overview of the metabolom by detecting as many metabolites as possible in the sample, thereby covering a broad range of metabolites from multiple metabolic pathways. If metabolites of potential interest are discovered, their fragmentation spectra are evaluated for identification. In a subsequent targeted analysis, the identity of the relevant metabolites can be confirmed and the results validated in larger sample sets.
A non-targeted metabolomics workflow is currently under development at the CECAD Lipidomics/Metabolomics Facility.
2. Shotgun Lipidomics
In the Shotgun Lipidomics approach, the total lipid extract is directly infused into the mass spectrometer, i.e. without chromatographic pre-separation. The mass spectrometric detection is accomplished by applying different scan modes, which are specific for the individual lipid classes. The Lipidomics/Metabolomics Facility uses the Shotgun Lipidomics approach for the targeted quantification of glycerolipids, glycerophospholipids, cholesteryl esters, and epidermal ceramides.
3. Gas Chromatography with Mass Selective Detection (GC/MSD)
Our GC/MSD system is mainly used for the determination of fatty acids after their derivatization to fatty acid methyl esters (FAMEs).
4. Analytical Thin Layer Chromatography (TLC)
TLC is a quick and robust method to get a general overview of the lipid composition of biological samples. Lipid classes differing from each other in their head groups (e.g. phosphatidylcholines and phosphatidylethanolamines) can be separated and quantified by densitometry. The Facility offers the determination of all common lipid classes by TLC.
Dr. Susanne Brodesser, Dipl. Chem.