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Updated: Mar 2, 2026

Capillary Electrophoresis-based Hydrogen/Deuterium Exchange for Conformational Characterization of Proteins with Top-down Mass Spectrometry
Published on: June 8, 2021
The use of deuterium oxide as a mobile phase for structural elucidation by HPLC/UV/ESI/MS
Olsen1, Cummings, Kennedy-Gabb
1Department of Analytical Sciences, SmithKline Beecham Pharmaceuticals, King of Prussia, Pennsylvania 19406, USA.
Abstract:
The use of deuterium oxide as a mobile phase in the routine analysis of pharmaceutical compounds was investigated. The deuterium exchange of labile hydrogen atoms aids in structural confirmation and elucidation of unknown impurities and degradation products. Although deuterium oxide as a mobile phase does in some cases change the retention times, the changes in retention times do not interfere with the analysis. A study of the high-performance liquid chromatography system shows that equilibration times for the deuterium-containing mobile phases are similar to equilibration times with changes of other mobile phases. The use of this technique in the analysis of pharmaceutical compounds and other small molecules is presented.
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