Glycans, carbohydrates, and glycoconjugates are involved in many crucial biological processes, such as disease development, immune responses, and cell–cell recognition. Glycans and carbohydrates are known for the large number of isomeric features associated with their structures, making analysis challenging compared with other biomolecules. Mass spectrometry has become the primary method of structural characterization for carbohydrates, glycans, and glycoconjugates. Metal adduction is especially important for the mass spectrometric analysis of carbohydrates and glycans. Metal‐ion adduction to carbohydrates and glycoconjugates affects ion formation and the three‐dimensional, gas‐phase structures. Herein, we discuss how metal‐ion adduction impacts ionization, ion mobility, ion activation and dissociation, and hydrogen/deuterium exchange for carbohydrates and glycoconjugates. We also compare the use of different metals for these various techniques and highlight the value in using metals as charge carriers for these analyses. Finally, we provide recommendations for selecting a metal for analysis of carbohydrate adducts and describe areas for continued research.

Electrospray ionization (ESI) is frequently used to produce gas-phase ions for mass spectrometry (MS)-based techniques. The composition of solvents used in ESI-MS is often manipulated to enhance analyte ionization, including for carbohydrates. Moreover, to characterize analyte structures, ESI has been coupled to hydrogen/deuterium exchange, ion mobility, and tandem MS. Therefore, it is important to understand how solvent composition affects the structure of carbohydrates during and after ESI. In this work, we use molecular dynamics to simulate the desolvation of ESI droplets containing a model carbohydrate and observe the formation of carbohydrate adducts with metal ions. Molecular-level details on the effects of formulating mixtures of water, methanol, and acetonitrile to achieve enhanced ionization are presented. We complement our simulations with ESI-MS experiments. We report that when sprayed from aqueous mixtures containing volatile solvents, carbohydrates ionize to form metal−ion adducts rapidly due to rapid solvent evaporation rather than changes in the ionization mechanism. We find that when sprayed from solvent mixtures, carbohydrates are primarily solvated by water due to the migration of more volatile solvents to the surface of the droplet. Ultimately, the structure of the carbohydrate varies depending on its solvent environment, as inter-and intramolecular interactions are affected. We propose that solvents with 25% or more water may be used to enhance the ionization of carbohydrates with minimal effect on the structure during and after ESI.