|
|
||||||||
Correspondence to:
G. Schmitz
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 C8-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 µg 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.Liebisch, G., W. Drobnik, M. Reil, B. Trümbach, R. Arnecke, B. Olgemöller, A. Roscher, and G. Schmitz. Quantitative measurement of different ceramide species from crude cellular extracts by electrospray ionization tandem mass spectrometry (ESI-MS/MS). J. Lipid Res. 1999. 40: 1539;1546.
Supplementary key words:
ceramide quantitation, electrospray ionization, tandem mass spectrometry
Copyright © 1999 by Lipid Research, Inc.
Methods
Quantitative measurement of different ceramide species from crude cellular extracts by electrospray ionization tandem mass spectrometry (ESI-MS/MS)
G. Liebischa,
W. Drobnika,
M. Reila,
B. Trümbacha,
R. Arneckeb,
B. Olgemöllerb,
A. Roscherc, and
G. Schmitza
a Institut für Klinische Chemie und Laboratoriumsmedizin, Klinikum der Universität Regensburg, Franz-Josef-Strauss-Allee 11, 93042 Regensburg, Germany
b Labor Becker, Olgemöller & Partner, München, Germany
c Kinderklinik, Klinische Chemie/Biochemie, Ludwig Maximilian Universität München, München, Germany
![]()
CiteULike
Complore
Connotea
Del.icio.us
Digg
Reddit
Technorati What's this?
This article has been cited by other articles:
![]() |
Y. Masukawa, H. Narita, E. Shimizu, N. Kondo, Y. Sugai, T. Oba, R. Homma, J. Ishikawa, Y. Takagi, T. Kitahara, et al. Characterization of overall ceramide species in human stratum corneum J. Lipid Res., July 1, 2008; 49(7): 1466 - 1476. [Abstract] [Full Text] [PDF] |
||||
![]() |
M. Kyogashima, K. Tadano-Aritomi, T. Aoyama, A. Yusa, Y. Goto, K. Tamiya-Koizumi, H. Ito, T. Murate, R. Kannagi, and A. Hara Chemical and Apoptotic Properties of Hydroxy-Ceramides Containing Long-Chain Bases with Unusual Alkyl Chain Lengths J. Biochem., July 1, 2008; 144(1): 95 - 106. [Abstract] [Full Text] [PDF] |
||||
![]() |
V. Matyash, G. Liebisch, T. V. Kurzchalia, A. Shevchenko, and D. Schwudke Lipid extraction by methyl-tert-butyl ether for high-throughput lipidomics J. Lipid Res., May 1, 2008; 49(5): 1137 - 1146. [Abstract] [Full Text] [PDF] |
||||
![]() |
M. Valsecchi, L. Mauri, R. Casellato, S. Prioni, N. Loberto, A. Prinetti, V. Chigorno, and S. Sonnino Ceramide and sphingomyelin species of fibroblasts and neurons in culture J. Lipid Res., February 1, 2007; 48(2): 417 - 424. [Abstract] [Full Text] [PDF] |
||||
![]() |
K. A. Walton, B. G. Gugiu, M. Thomas, R. J. Basseri, D. R. Eliav, R. G. Salomon, and J. A. Berliner A role for neutral sphingomyelinase activation in the inhibition of LPS action by phospholipid oxidation products J. Lipid Res., September 1, 2006; 47(9): 1967 - 1974. [Abstract] [Full Text] [PDF] |
||||
![]() |
Y. Masukawa, H. Tsujimura, and H. Narita Liquid chromatography-mass spectrometry for comprehensive profiling of ceramide molecules in human hair J. Lipid Res., July 1, 2006; 47(7): 1559 - 1571. [Abstract] [Full Text] [PDF] |
||||
![]() |
S. C.D. van IJzendoorn, J. M. van der Wouden, G. Liebisch, G. Schmitz, and D. Hoekstra Polarized Membrane Traffic and Cell Polarity Development Is Dependent on Dihydroceramide Synthase-Regulated Sphinganine Turnover Mol. Biol. Cell, September 1, 2004; 15(9): 4115 - 4124. [Abstract] [Full Text] [PDF] |
||||
![]() |
T. Herrmann, F. van der Hoeven, H.-J. Grone, A. F. Stewart, L. Langbein, I. Kaiser, G. Liebisch, I. Gosch, F. Buchkremer, W. Drobnik, et al. Mice with targeted disruption of the fatty acid transport protein 4 (Fatp 4, Slc27a4) gene show features of lethal restrictive dermopathy J. Cell Biol., June 23, 2003; 161(6): 1105 - 1115. [Abstract] [Full Text] [PDF] |
||||
![]() |
W. Drobnik, G. Liebisch, F.-X. Audebert, D. Frohlich, T. Gluck, P. Vogel, G. Rothe, and G. Schmitz Plasma ceramide and lysophosphatidylcholine inversely correlate with mortality in sepsis patients J. Lipid Res., April 1, 2003; 44(4): 754 - 761. [Abstract] [Full Text] [PDF] |
||||
![]() |
R. Sandhoff, S. T. Hepbildikler, R. Jennemann, R. Geyer, V. Gieselmann, R. L. Proia, H. Wiegandt, and H.-J. Grone Kidney Sulfatides in Mouse Models of Inherited Glycosphingolipid Disorders. DETERMINATION BY NANO-ELECTROSPRAY IONIZATION TANDEM MASS SPECTROMETRY J. Biol. Chem., May 31, 2002; 277(23): 20386 - 20398. [Abstract] [Full Text] [PDF] |
||||
![]() |
T. Murate, M. Suzuki, M. Hattori, A. Takagi, T. Kojima, T. Tanizawa, H. Asano, T. Hotta, H. Saito, S. Yoshida, et al. Up-regulation of Acid Sphingomyelinase during Retinoic Acid-induced Myeloid Differentiation of NB4, a Human Acute Promyelocytic Leukemia Cell Line J. Biol. Chem., March 15, 2002; 277(12): 9936 - 9943. [Abstract] [Full Text] [PDF] |
||||
![]() |
C. Vieu, F. Terce, F. Chevy, C. Rolland, R. Barbaras, H. Chap, C. Wolf, B. Perret, and X. Collet Coupled assay of sphingomyelin and ceramide molecular species by gas liquid chromatography J. Lipid Res., March 1, 2002; 43(3): 510 - 522. [Abstract] [Full Text] [PDF] |
||||
![]() |
T. Kanto, P. Kalinski, O. C. Hunter, M. T. Lotze, and A. A. Amoscato2 Ceramide Mediates Tumor-Induced Dendritic Cell Apoptosis J. Immunol., October 1, 2001; 167(7): 3773 - 3784. [Abstract] [Full Text] [PDF] |
||||
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
| All ASBMB Journals | Journal of Biological Chemistry |
| Molecular and Cellular Proteomics | ASBMB Today |