High-Performance Liquid Chromatography Analysis of Ganglioside Carbohydrates at the Picomole Level after CERAMIDE GLYCANASE (cas 106769-66-8) Digestion and Fluorescent Labeling with 2-Aminobenzamide

Add time:08/09/2019    Source:sciencedirect.com

The functional importance of glycolipids has emphasized the need for more sensitive methods of detection, characterization, and quantification than has often been possible using traditional thin-layer chromatographic techniques. We describe the use of CERAMIDE GLYCANASE (cas 106769-66-8) and HPLC to identify and quantify gangliosides in which the carbohydrate is in Glcβ1→ linkage with ceramide. Detection of released carbohydrate was by fluorescent labeling with 2-aminobenzamide at the reducing terminal prior to HPLC analysis. Under the conditions described, ceramide glycanase hydrolyzed all of the common gangliosides studied, offering a broad spectrum of specificity. Release and detection of carbohydrate were linear over a wide range (over two orders of magnitude) of micromolar glycolipid substrate concentrations. Use of an N-linked glycan as an internal standard allowed accurate quantification and a recovery of 93% was achieved. The method additionally maintained the sensitivity (chromatographic peaks containing 1 pmol were readily detected from tissue samples) and comparable resolution to related assays. This was shown by the separation, not only of isomeric carbohydrates from the “a” and “b” series, but also of ganglioside carbohydrate differing only by the presence of either N-acetyl- or N-glycolylneuraminic acid. Application of the method to neutral glycosphingolipids and to tissue samples, including 10-μl quantities of plasma, is illustrated. Glycan structures were confirmed by exoglycosidase digestion and/or matrix-assisted laser desorption/ionization mass spectrometry.

We also recommend Trading Suppliers and Manufacturers of CERAMIDE GLYCANASE (cas 106769-66-8). Pls Click Website Link as below: cas 106769-66-8 suppliers

Prev:A unique glycosphingolipid-splitting enzyme (ceramide-glycanase from leech) cleaves the linkage between the oligosaccharide and the ceramide
Next:Synthesis of aryl 3′-sulfo-β-lactosides as fluorogenic and chromogenic substrates for CERAMIDE GLYCANASE (cas 106769-66-8)s)