ORBi: Fillet Marianne - Separation, identification and quantitation of ceramides in human cancer cells by liquid chromatography–electrospray ionization tandem mass spectrometry

Reference : Separation, identification and quantitation of ceramides in human cancer cells by liquid...


	Document type :	Scientific journals : Article
	Discipline(s) :	Human health sciences : Pharmacy, pharmacology & toxicology
	To cite this reference:	http://hdl.handle.net/2268/13395

Title :	Separation, identification and quantitation of ceramides in human cancer cells by liquid chromatography–electrospray ionization tandem mass spectrometry
Language :	English
Author, co-author :	Fillet, Marianne [Université de Liège - ULg > Département de pharmacie > Analyse des médicaments >]
	Van Heugen, Jean-Claude [Centre Hospitalier Universitaire de Liège - CHU > > Chimie médicale >]
	Servais, Anne-Catherine [Université de Liège - ULg > Département de pharmacie > Analyse des médicaments >]
	De Graeve, Jean [Université de Liège - ULg > Services généraux (Faculté de médecine) > Relations académiques et scientifiques (Médecine) >]
	Crommen, Jacques [Université de Liège - ULg > Département de pharmacie > Analyse des médicaments >]
Publication date :	2002
Journal title :	Journal of Chromatography. A
Publisher :	Elsevier Science
Volume :	949
Pages :	225-233
Audience :	International
ISSN :	0021-9673
City :	Amsterdam
Country :	The Netherlands
Keywords :	[en] 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 units :	Centre Interfacultaire de Recherches Bioanalytiques et Biopharmaceutiques - CRBB
Funders :	Fonds de la Recherche Scientifique (Communauté française de Belgique) - F.R.S.-FNRS
Target :	Researchers ; Professionals
Permalink :	http://hdl.handle.net/2268/13395

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Open access	LCMSceramideJChromA2002.pdf		Publisher postprint	184.99 kB	View/Open

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