Article (Scientific journals)
Separation, identification and quantitation of ceramides in human cancer cells by liquid chromatography–electrospray ionization tandem mass spectrometry
Fillet, Marianne; Van Heugen, Jean-Claude; Servais, Anne-Catherine et al.
2002In Journal of Chromatography. A, 949, p. 225-233
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Keywords :
Ceramides; Sphingolipids; Lipids
Abstract :
[en] Ceramides are important intracellular second messengers that play a role in the regulation of cell growth, differentiation and programmed cell death. Qualitative and quantitative analysis of these second messengers requires sensitive and specific analytical methods to detect endogenous levels of individual ceramide species and to differentiate between them. Nine synthetic ceramides were separated by liquid chromatography coupled to tandem mass spectrometry on a C bonded silica 18 column. The lipids were eluted in gradient elution mode using a mixture of water, acetonitrile and 2-propanol as mobile phase. They were detected by reaction monitoring performed on positive ion electrospray generated ions. Collision-induced fragmentations conducted on ceramides produced a well characteristic product ion at m/z 264, making multiple reaction monitoring (MRM) well suited for various ceramides quantitative measurements. After optimization of the extraction step, the proposed methodology was able to identify and quantify different ceramide species issued from human cancer cells. The method could be validated for C , C and C ceramides, quantified at the nanogram level. The validation exhibits good 16 18 20 results with respect to linearity, accuracy and precision.
Research center :
CRBB - Centre Interfacultaire de Recherches Bioanalytiques et Biopharmaceutiques - ULiège
Disciplines :
Pharmacy, pharmacology & toxicology
Author, co-author :
Fillet, Marianne ;  Université de Liège - ULiège > Département de pharmacie > Analyse des médicaments
Van Heugen, Jean-Claude ;  Centre Hospitalier Universitaire de Liège - CHU > Chimie médicale
Servais, Anne-Catherine  ;  Université de Liège - ULiège > Département de pharmacie > Analyse des médicaments
De Graeve, Jean ;  Université de Liège - ULiège > Services généraux (Faculté de médecine) > Relations académiques et scientifiques (Médecine)
Crommen, Jacques ;  Université de Liège - ULiège > Département de pharmacie > Analyse des médicaments
Language :
English
Title :
Separation, identification and quantitation of ceramides in human cancer cells by liquid chromatography–electrospray ionization tandem mass spectrometry
Publication date :
2002
Journal title :
Journal of Chromatography. A
ISSN :
0021-9673
eISSN :
1873-3778
Publisher :
Elsevier Science, Amsterdam, Netherlands
Volume :
949
Pages :
225-233
Peer reviewed :
Peer Reviewed verified by ORBi
Funders :
F.R.S.-FNRS - Fonds de la Recherche Scientifique [BE]
Available on ORBi :
since 28 May 2009

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