The usefulness of hydrazine derivatives for mass spectrometric analysis of carbohydrates

Abstract

Over the last years, extensive studies have evaluated glycans from different biological samples and validated the importance of glycosylation as one of the most important post-translational modifications of proteins. Although a number of new methods for carbohydrate analysis have been published and there has been significant progress in their identification, the development of new approaches to study these biomolecules and understand their role in living systems are still vivid challenges that intrigue glycobiologists. In the last decade, the success in analyses of oligosaccharides has been driven mainly by the development of innovative, highly sensitive mass spectrometry techniques. For enhanced mass spectrometry detection, carbohydrate molecules are often derivatized. Besides, the type of labeling can influence the fragmentation pattern and make the structural analysis less complicated. In this regard, in 2003 we introduced the low scale, simple non-reductive tagging of glycans employing phenylhydrazine (PHN) as the derivatizing reagent. PHN-labeled glycans showed increased detection and as reported previously they can be analyzed by HPLC, ESI, or MALDI immediately after derivatization. Under tandem mass spectrometry conditions, PHN-derivatives produced useful data for the structural elucidation of oligosaccharides. This approach of analysis has helped to reveal new isomeric structures for glycans of known/unknown composition and has been successfully applied for the profiling of N-glycans obtained from serum samples and cancer cells. The efficacy of this labeling has also been evaluated for different substituted hydrazine reagents. This review summarizes all types of reducing-end labeling based on hydrazone-linkage that have been used for mass spectrometric analyses of oligosaccharides. This review is also aimed at correcting some past misconceptions or interpretations reported in the literature.

N-Glycome Profiling of Patatins from Different Potato Species of Solanum Genus

“…23,28 In the spectra of glycoproteins isolated from species Solanum andigenum and Solanum stenotomum, additional peaks appeared at m/z 1671.6 and 1833.7 (Figure 1c). Tandem mass spectra showed that these N-glycans had both antennae occupied with hexose residues only, and the fragmentation pattern was similar to that assigned to high-mannose structures as previously described.…”

Analysis of carbohydrates and glycoconjugates by matrix‐assisted laser desorption/ionization mass spectrometry: An update for 2013–2014

“…A review on the use of hydrazine derivatives for the mass spectrometry of carbohydrates has been written by Lattová and Perreault () (104 cited references). Derivatization of carbohydrates as hydrazides ( 49 ) has been achieved in as little as 5 min.…”

Purification of Derivatized Oligosaccharides by Solid Phase Extraction for Glycomic Analysis

“…Thus, the analysis of glycosylation is extensively conducted in biological and clinical research as well as in pharmaceutical industry [8]–[10]. Different methods have been reported, including high performance liquid chromatography (HPLC) [11], [12], capillary electrophoresis [13], [14] and high pH anion-exchange chromatography [15], [16] with electrochemical, optical or mass spectrometric (MS) detection [17], [18]. To enhance the detection sensitivity, oligosaccharides are usually derivatized prior to analysis.…”

“…To enhance the detection sensitivity, oligosaccharides are usually derivatized prior to analysis. Most of such derivatization reactions can be accomplished by coupling the reagents with the reducing end of saccharides [17], [19]. During this step, reagents, including salts, derivatization reagents and solvent are normally presented in large excess.…”