Abstract
High resolution and tandem mass spectrometry are popular techniques for the structure elucidation of small molecule impurities and metabolites. However, proposing definitive structures for isomers solely based on the mass spectrometry data can be challenging. Hydrogen/Deuterium exchange mass spectrometry can be used as an effective tool for the structure elucidation of isomers by determining the number of exchangeable hydrogen atoms on heteroatomic functional groups. The study demonstrates a simple and effective online hydrogen/deuterium exchange technique using two-dimensional liquid chromatography mass spectrometry for unknown impurity and metabolite structure elucidation. A mixture of isomeric compounds were separated in the first dimension of the two-dimensional liquid chromatography and transferred to the second dimension using heart-cutting valve. The hydrogen/deuterium exchange was carried out in the second dimension using deuterium oxide as one of the mobile phase solvents. The eluents were analyzed after hydrogen/deuterium exchange with mass spectrometry. The application of two-dimensional liquid chromatography mass spectrometry for hydrogen/deuterium exchange experiments provides unique advantages of simplicity along with minimized time and resources.
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Acknowledgements
We would like to thank Daniel Knueppel, Mingming Ma, and Jeff Gilbert for their guidance and support for this project.
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This work was supported by Corteva Agriscience.
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Johnson, J.T., Patil, R.A. & Zu, C. Two-Dimensional Liquid Chromatography–Mass Spectrometry: Heart-Cutting Hydrogen/Deuterium Exchange as a Tool for Small Molecule Structure Elucidation. Chromatographia 86, 535–540 (2023). https://doi.org/10.1007/s10337-023-04265-0
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DOI: https://doi.org/10.1007/s10337-023-04265-0