Quantitative analysis of phospholipids containing arachidonate and docosahexaenoate chains in microdissected regions of mouse brain
| Autor: | Robert C. Murphy, Paul H. Axelsen |
|---|---|
| Rok vydání: | 2009 |
| Předmět: |
Plasmalogen
Docosahexaenoic Acids Phospholipid normal-phase HPLC plasmalogen QD415-436 Chemical Fractionation Biochemistry Mass Spectrometry chemistry.chemical_compound Mice Endocrinology Alzheimer Disease Methods Animals Phospholipids Chromatography Arachidonic Acid negative-mode MS Selected reaction monitoring Brain Cell Biology Phosphatidic acid Oxidative Stress chemistry Docosahexaenoic acid Glycerophospholipid Arachidonic acid lipids (amino acids peptides and proteins) Female Quantitative analysis (chemistry) Alzheimer’s disease Microdissection |
| Zdroj: | Journal of Lipid Research, Vol 51, Iss 3, Pp 660-671 (2010) |
| ISSN: | 1539-7262 |
| Popis: | Phospholipids containing polyunsaturated fatty acyl chains are prevalent among brain lipids, and regional differences in acyl chain distribution appear to have both functional and pathological significance. A method is described in which the combined application of GC and multiple reaction monitoring (MRM) MS yielded precise relative quantitation and approximate absolute quantitation of lipid species containing a particular fatty acyl chain in milligram-sized tissue samples. The method uses targeted MRM to identify specific molecular species of glycerophosphocholine lipids, glycerophospho-ethanolamine lipids, glycerophosphoinositol lipids, glycerophosphoserine lipids, glycero-phosphoglycerol lipids, and phosphatidic acids that contain esterified arachidonate (AA) and docosahexaenoate (DHA) separated during normal phase LC/MS/MS analysis. Quantitative analysis of the AA and DHA in the LC fractions is carried out using negative ion chemical ionization GC/MS and stable isotope dilution strategies. The method has been applied to assess the glycerophospholipid molecular species containing AA and DHA in microdissected samples of murine cerebral cortex and hippocampus. Results demonstrate the potential of this approach to identify regional differences in phospholipid concentration and reveal differences in specific phospholipid species between cortex and hippocampus. These differences may be related to the differential susceptibility of different brain regions to neurodegenerative disorders. |
| Databáze: | OpenAIRE |
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