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Updated: Aug 13, 2026

Capillary Electrophoresis-based Hydrogen/Deuterium Exchange for Conformational Characterization of Proteins with Top-down Mass Spectrometry
Published on: June 8, 2021
Ultra performance liquid chromatography (UPLC) further improves hydrogen/deuterium exchange mass spectrometry
Yan Wu1, John R Engen, William B Hobbins
1Department of Chemistry, University of New Mexico, Albuquerque, New Mexico 87131, USA.
Abstract:
Ultra performance liquid chromatography (UPLC) employs particles smaller than 2 microm in diameter to achieve superior resolution, speed, and sensitivity compared with high-performance liquid chromatography (HPLC). We have tested the suitability of UPLC for the analysis of deuterated peptides in hydrogen exchange mass spectrometry experiments. Superior resolution and sample throughput were obtained with UPLC versus HPLC. For highly deuterated model peptides, deuterium loss using UPLC was greater than the deuterium loss observed using a conventional HPLC system, primarily as a result of the injection requirements of the UPLC system. Partially deuterated cytochrome c peptides also lost more deuterium in UPLC versus HPLC, although the effect was not as pronounced as it was for the highly deuterated model peptides. The exceptional chromatographic aspects of UPLC make it a very attractive alternative to HPLC for hydrogen exchange mass spectrometry experiments.
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