HPLC separation and ultrasensitive optical quantification of ceramide species applying 7-(diethylamino)coumarin-3-carbonyl azide derivatisation

Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences
Markus BlaessHans-Peter Deigner

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

Feb 1, 1993·Comparative Biochemistry and Physiology. B, Comparative Biochemistry·S O ChanD C Yeung
Oct 16, 1999·The Journal of Biological Chemistry·R L ThomasA A Amoscato
Jan 5, 2002·Analytical Biochemistry·A BielawskaY A Hannun
Aug 1, 1959·Canadian Journal of Biochemistry and Physiology·E G BLIGH, W J DYER
May 18, 2004·RNA·Xinliang Zhao, Yi-Tao Yu
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

❮ Previous
Next ❯

Related Concepts

Related Feeds

Anxiety Disorders

Discover the latest research on anxiety disorders including agoraphobia, panic disorder, obsessive-compulsive disorder, and post-traumatic stress disorder here.

Related Papers

Hormone and Metabolic Research = Hormon- Und Stoffwechselforschung = Hormones Et Métabolisme
M GórskaZ Namiot
Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences
Bart A J van LandeghemHenk A Claessens
Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences
Jie ChenMichael J Bennett
© 2022 Meta ULC. All rights reserved