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HPLC Separation and Ultrasensitive Optical Quantification of Ceramide Species Applying 7-(diethylamino)coumarin-3-carbonyl Azide Derivatisation

HPLC Separation and Ultrasensitive Optical Quantification of Ceramide Species Applying 7-(diethylamino)coumarin-3-carbonyl Azide Derivatisation

Markus Blaess, Ralf A Claus, Hans-Peter Deigner

J Chromatogr B Analyt Technol Biomed Life Sci. 2015 Apr 1;986-987:123-8.

PMID: 25728369

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.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
AP157673160	7-(Diethylamino)coumarin-3-carbonyl azide		157673-16-0	Price
AP85985440	9-Anthracenecarbonyl cyanide		85985-44-0	Price

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