'Hypermethylation' of anthranilic acid-labeled sugars confers the selectivity required for liquid chromatography-mass spectrometry
| Autor: | Elisabeth Svehla, Réka Mócsai, Rudolf Figl, Paul Kosma, Friedrich Altmann, Markus Windwarder, Erika Staudacher, Jean-Baptiste Farcet |
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| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Spectrometry Mass Electrospray Ionization Iodide Biophysics Carbohydrates Mass spectrometry Biochemistry Methylation Online Systems Fluorescence 03 medical and health sciences chemistry.chemical_compound Hydrolysis Liquid chromatography–mass spectrometry Tandem Mass Spectrometry Anthranilic acid Organic chemistry Monosaccharide ortho-Aminobenzoates Molecular Biology Chromatography High Pressure Liquid chemistry.chemical_classification Chromatography Reverse-Phase Chromatography Monosaccharides Cell Biology 030104 developmental biology chemistry Solvents Selectivity |
| Zdroj: | Analytical biochemistry. 514 |
| ISSN: | 1096-0309 |
| Popis: | Analysis of the monosaccharides of complex carbohydrates is often performed by liquid chromatography with fluorescence detection. Unfortunately, methylated sugars, unusual amino- or deoxysugars and incomplete hydrolysis can lead to erroneous assignments of peaks. Here, we demonstrate that a volatile buffer system is suitable for the separation of anthranilic acid labeled sugars. It allows off-line examination of peaks by electrospray mass spectrometry. Approaches towards on-line mass spectrometric detection using reversed-phase or porous graphitic carbon columns fell short of achieving sufficient separation of the relevant isobaric sugars. Adequate chromatographic performance for isomeric sugars was achieved with reversed-phase chromatography of “hyper”-methylated anthranilic acid-labeled monosaccharides. Deuteromethyl iodide facilitates the discovery of naturally methylated sugars and identification of their parent monosaccharide as demonstrated with N-glycans of the snail Achatina fulica, where two thirds of the galactoses and a quarter of the mannoses were methylated. |
| Databáze: | OpenAIRE |
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