In-ESI H/D exchange is a convenient technique for analyzing small-molecular complex mixtures. However, such experiments do not yield sufficient levels of exchange or require an elevated temperature of the ion transfer capillary. Increased temperature may result in unexpected additional exchanges of -CH groups that may complicate the interpretation of the H/D exchange data used for identification. Gas-phase H/D exchange depends on the gas-phase basicity of the deuterating agent. In-ESI exchange involves both droplet-phase and gas-phase mechanisms, depending on a particular ion source setup and the deuterating agent used. Therefore, the addition of strong bases to the reaction mixture should facilitate in-ESI exchange. This work aimed to investigate the capabilities of different amines to improve in-ESI H/D exchange compared with pure D2O and to choose an amine modifier to increase the extent of H/D exchange. It was shown that such additives substantially enhanced the extent of H/D exchange in small molecules, peptides, and proteins even without heating the capillary. It was found that the extent of exchange increases in the following order: tertiary amines < secondary amines < primary amines. Therefore, we suggest that amines act as deuterating agents after being exchanged with D2O. These findings may improve H/D exchange applications, especially in small molecule analysis. The observation of improved H/D exchange with amine additives in peptides and proteins may become a subject of future research.