Liebisch, Gerhard and Drobnik, W. and Reil, M. and Truembach, B. and Arnecke, Ralf and Olgemoeller, B. and Roscher, A. and Schmitz, Gerd (1999) Quantitative measurement of different ceramide species from crude cellular extracts by electrospray ionization tandem mass spectrometry (ESI-MS/MS). JOURNAL OF LIPID RESEARCH, 40 (8). pp. 1539-1546. ISSN 0022-2275,
Full text not available from this repository. (Request a copy)Abstract
Ceramide (CER) is an important signaling molecule involved in a variety of cellular processes, including differentiation, cell growth, and apoptosis, Currently, different techniques are applied for CER quantitation, some of which are relatively insensitive and/or time consuming, Tandem mass spectrometry with its high selectivity and sensitivity is a very useful technique for detection of low abundant metabolites without prior purification or derivatization, In contrast to existing mass spectrometry methods, the developed electrospray tandem mass spectrometry (ESI-MS/MS) technique is capable of quantifying different CER species from crude cellular lipid extracts. The ESI-MS/MS is performed with a continuous flow injection and the use of an autosampler, resulting in a high throughput capability, The collision-induced fragmentation of CER produced, in addition to others, a characteristic fragment of m/z 264, making a precursor ion scan of 264 well suited for CER quantitation. Quantitation is achieved by use of a constant concentration of a non-naturally occurring internal standard CS-CER, together with a calibration curve established by spiking different concentrations of naturally occurring CER, The calibration curves showed linearity over a wide concentration range and sample volumes equivalent to 10 mu g of cell protein corresponding to about 20,000 fibroblasts were sufficient for CER analysis, Moreover this assay showed a detection limit at the subpicomole level. In summary, this methodology enables accurate and rapid analysis of CER from small samples without prior separation steps, thus providing a useful tool for signal transduction research.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | PERFORMANCE LIQUID-CHROMATOGRAPHY; INDUCED APOPTOSIS; SPHINGOMYELIN PATHWAY; CELLS; SPHINGOSINE; SYNTHASE; OCCURS; ROLES; ceramide quantitation; electrospray ionization; tandem mass spectrometry |
| Subjects: | 600 Technology > 610 Medical sciences Medicine |
| Divisions: | Medicine > Lehrstuhl für Klinische Chemie und Laboratoriumsmedizin |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 11 Jan 2023 13:46 |
| Last Modified: | 11 Jan 2023 13:46 |
| URI: | https://pred.uni-regensburg.de/id/eprint/49091 |
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