Labelling saccharides with phenylhydrazine for electrospray and matrix-assisted laser desorption–ionization mass spectrometry

• Published in: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences Vol. 793; no. 1; pp. 167 - 179
• Main Authors: Lattova, Erika, Perreault, Hélène
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 05.08.2003
• Subjects: Carbohydrates - chemistry; Chromatography, High Pressure Liquid - methods; Glycosylation; Phenylhydrazines - chemistry; Phenylhydrazone; Saccharides; Spectrometry, Mass, Electrospray Ionization - methods; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization - methods; Phenylhydrazone; Saccharides
• ISSN: 1570-0232; 1873-376X
• DOI: 10.1016/S1570-0232(03)00374-X

A well-known reaction of carbonyl compounds with phenylhydrazine has been applied to saccharides, providing increased sensitivity for mass spectrometric (MS) and ultraviolet (UV) detection during high-performance liquid chromatographic (HPLC) separations. After a simple derivatization procedure for 1 h at 70 °C and purification of the reaction mixture from excess reagent by extraction, the sugar derivatives were characterized by direct injection or on-line HPLC/electrospray ionization (ESI) and by matrix-assisted laser desorption/ionization (MALDI) MS. Because no salts are used or produced upon reaction, this procedure is very simple and suitable for the tagging of saccharides. The reaction allows for on-target derivatization and products are very stable. The derivatization procedure has been applied to commercially-obtained small saccharides and standard N-linked oligosaccharides. Lastly, hen ovalbumin N-glycans were detached enzymatically and characterized by MALDI–MS as their phenylhydrazone derivatives.