Scherer, Max and Leuthaeuser-Jaschinski, Kerstin and Ecker, Josef and Schmitz, Gerd and Liebisch, Gerhard (2010) A rapid and quantitative LC-MS/MS method to profile sphingolipids. JOURNAL OF LIPID RESEARCH, 51 (7). pp. 2001-2011. ISSN 0022-2275, 1539-7262
Full text not available from this repository. (Request a copy)Abstract
Sphingolipids comprise a highly diverse and complex class of molecules that serve not only as structural components of membranes but also as signaling molecules. To understand the differential role of sphingolipids in a regulatory network, it is important to use specific and quantitative methods. We developed a novel LC-MS/MS method for the rapid, simultaneous quantification of sphingolipid metabolites, including sphingosine, sphinganine, phyto-sphingosine, di- and trimethyl-sphingosine, sphingosylphosphorylcholine, hexosylceramide, lactosylceramide, ceramide-1-phosphate, and dihydroceramide-1-phosphate. Appropriate internal standards (ISs) were added prior to lipid extraction. In contrast to most published methods based on reversed phase chromatography, we used hydrophilic interaction liquid chromatography and achieved good peak shapes, a short analysis time of 4.5 min, and, most importantly, coelution of analytes and their respective ISs. To avoid an overestimation of species concentrations, peak areas were corrected regarding isotopic overlap where necessary. Quantification was achieved by standard addition of naturally occurring sphingolipid species to the sample matrix. The method showed excellent precision, accuracy, detection limits, and robustness. As an example, sphingolipid species were quantified in fibroblasts treated with myriocin or sphingosine-kinase inhibitor.jlr In summary, this method represents a valuable tool to evaluate the role of sphingolipids in the regulation of cell functions.-Scherer, M., K. Leuthauser-Jaschinski, J. Ecker, G. Schmitz, and G. Liebisch. A rapid and quantitative LC-MS/MS method to profile sphingolipids. J. Lipid Res. 2010. 51: 2001-2011
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | TANDEM MASS-SPECTROMETRY; HIGH-THROUGHPUT QUANTIFICATION; SPHINGOSINE 1-PHOSPHATE; LYSOPHOSPHATIDIC ACID; CELLULAR-EXTRACTS; PLASMA SAMPLES; ESI-MS/MS; SPHINGOSINE-1-PHOSPHATE; ATHEROSCLEROSIS; SPHINGANINE; HILIC; high throughput; ESI; free sphingoid base; methylated sphingoid base; hexosylceramide; lactosylceramide; sphingosylphosphorylcholine; ceramide-1-phosphate |
| Subjects: | 600 Technology > 610 Medical sciences Medicine |
| Divisions: | Medicine > Lehrstuhl für Klinische Chemie und Laboratoriumsmedizin |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 27 Jul 2020 11:45 |
| Last Modified: | 27 Jul 2020 11:45 |
| URI: | https://pred.uni-regensburg.de/id/eprint/24516 |
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