Abstract
Ceramides are derivatised using 7-(diethylamino)coumarin-3-carbonyl azide; subsequent gradient HPLC separation allows sensitive optical quantification of individual cellular ceramides. Compared to 9-anthracenecarbonyl cyanide (9-anthroyl nitrile) as derivatisation agent, the limit of detection could be improved 415-fold, respectively 10,000-fold (detection limit 0.6 pmol labelled ceramide/sample) when compared to benzoyl chloride-labelling. Acidic or alkaline catalysts are not required, enabling drying and storing of the labelled samples and a free choice of solvents for subsequent HPLC-separation. The quantitative method is characterised by high sensitivity, linearity and robustness in the pico- to nanomolar concentration range and does not require mass-spectrometry for quantification of cellular ceramides.
References
Jul 29, 1987·Journal of Chromatography·S SnadaY Nagai
Apr 29, 1994·Journal of Chromatography. a·K KamataM Nishijima
Dec 1, 1993·Archives of Biochemistry and Biophysics·R BruP L Luisi
Feb 1, 1993·Comparative Biochemistry and Physiology. B, Comparative Biochemistry·S O ChanD C Yeung
Jan 15, 1996·Analytical Biochemistry·C BalestrieriL Servillo
Jan 15, 1997·Analytical Biochemistry·M GuR Aebersold
Oct 16, 1999·The Journal of Biological Chemistry·R L ThomasA A Amoscato
Jan 5, 2002·Analytical Biochemistry·A BielawskaY A Hannun
Mar 7, 2002·Analytical Biochemistry·Xianlin Han
Aug 1, 1959·Canadian Journal of Biochemistry and Physiology·E G BLIGH, W J DYER
May 18, 2004·RNA·Xinliang Zhao, Yi-Tao Yu
Oct 4, 2008·Journal of Lipid Research·Philipp WiesnerGerhard Liebisch
Jun 13, 2009·Current Medicinal Chemistry·Ralf A ClausH Peter Deigner
Jun 29, 2011·Anti-cancer Agents in Medicinal Chemistry·Carmen BediaPatrice Codogno
Dec 3, 2011·Progress in Lipid Research·Sabine GröschGerd Geisslinger
Nov 16, 2012·Journal of Lipid Research·Megan M YoungHong-Gang Wang
Citations
Jun 7, 2016·Journal of Chromatography. a·Xumei LvShouzhuo Yao
Jun 25, 2016·Molecular Medicine·Ha-Yeun ChungRalf A Claus
Sep 5, 2019·Methods and Protocols·Srinath PashikantiJared J Barrott
Jan 1, 2015·Cell Death Discovery·M BlaessH-P Deigner