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Updated: Aug 12, 2026

Time-resolved ElectroSpray Ionization Hydrogen-deuterium Exchange Mass Spectrometry for Studying Protein Structure and Dynamics
Published on: April 17, 2017
Liquid chromatography-mass spectrometry using the hydrogen/deuterium exchange reaction as a tool for impurity
1Merck Research Laboratories, Analytical Research Department, P.O. Box 2000, RY 818-B112, Rahway, NJ 07065, USA. thomas_novak@merck.com
Abstract:
HPLC-MS employing deuterium oxide and common MS-compatible deuterated additives in the mobile phase with electrospray ionization is shown to be a viable approach for the structural elucidation of impurities in pharmaceutically active agents following initial studies with protic mobile phases. This approach incorporates the hydrogen/deuterium (H/D) exchange reaction where deuterium is substituted for hydrogen at labile sites. Some developmental compounds studied include an amide, amine, lipopeptide, indole and methyl sulfone. H/D exchange is rapid and the chromatographic performance using deuterated mobile phases is comparable to protic counterparts.
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