Abstract
Diacylglycerol (DAG) is an important intermediate of lipid metabolism and a component of phospholipase C signal transduction. Quantification of DAG in plant membranes represents a challenging task because of its low abundance. DAG can be measured by direct infusion mass spectrometry (MS) on a quadrupole time-of-flight mass spectrometer after purification from the crude plant lipid extract via solid-phase extraction on silica columns. Different internal standards are employed to compensate for the dependence of the MS and MS/MS signals on the chain length and the presence of double bonds in the acyl moieties. Thus, using a combination of single MS and MS/MS experiments, quantitative results for the different molecular species of DAGs from Arabidopsis can be obtained.
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Acknowledgments
We would like to thank Helga Peisker (University of Bonn) for help during sample preparation, method development and data evaluation. This work was supported by an Instrument Grant (Forschungsgrossgeräte-Antrag) and by the Sonderforschungsbereich 645 of Deutsche Forschungsgemeinschaft.
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vom Dorp, K., Dombrink, I., Dörmann, P. (2013). Quantification of Diacylglycerol by Mass Spectrometry. In: Munnik, T., Heilmann, I. (eds) Plant Lipid Signaling Protocols. Methods in Molecular Biology, vol 1009. Humana, Totowa, NJ. https://doi.org/10.1007/978-1-62703-401-2_5
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DOI: https://doi.org/10.1007/978-1-62703-401-2_5
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