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Updated: Jul 10, 2026

Capillary Electrophoresis-based Hydrogen/Deuterium Exchange for Conformational Characterization of Proteins with Top-down Mass Spectrometry

Capillary Electrophoresis-based Hydrogen/Deuterium Exchange for Conformational Characterization of Proteins with Top-down Mass Spectrometry

Published on: June 8, 2021

Reliable electrophoretic mobilities free from Joule heating effects using CE.

Christopher J Evenhuis¹, Vlastimil Hruska, Rosanne M Guijt

¹Australian Centre for Research on Separation Science, University of Tasmania, School of Chemistry, Hobart, Tasmania, Australia.

Electrophoresis

|October 18, 2007

Summary

This study introduces a method to accurately measure electrophoretic mobility in capillary electrophoresis (CE) by extrapolating to zero power dissipation, correcting for Joule heating effects. This approach significantly improves agreement with literature values for ionic mobility.

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Area of Science:

Analytical Chemistry
Physical Chemistry
Separation Science

Background:

Electrophoretic mobility values from capillary electrophoresis (CE) experiments often differ from accepted values derived from ionic conductance measurements.
The influence of Joule heating, a consequence of electrical current in electrolytes, on electrophoretic mobility has been historically underestimated.
Ionic strength significantly affects electrophoretic mobility, a factor well-documented in electrochemical studies.

Purpose of the Study:

To develop a straightforward and reproducible method for determining accurate electrophoretic mobilities using CE.
To mitigate the overestimation of mobility caused by Joule heating during CE experiments.
To enhance the agreement between experimentally determined and literature values of limiting ionic mobility.

Main Methods:

Electrophoretic mobility measurements were conducted across a spectrum of driving powers.
Extrapolation to zero power dissipation was employed to eliminate the impact of Joule heating.
The freeware program PeakMaster 5 was utilized for ionic strength corrections to calculate limiting ionic mobilities.

Main Results:

The developed method significantly improved the agreement between experimental and literature values of limiting ionic mobility by over an order of magnitude.
For inorganic anions in Tris-chromate BGE, mobility values increased by an average of 12.6% due to the elimination of Joule heating effects.
Ionic strength corrections reduced mobilities by 11% for univalent and 28% for divalent inorganic ions relative to their limiting values.

Conclusions:

The extrapolation to zero power dissipation is a reliable technique for obtaining accurate electrophoretic mobilities in CE.
Accurate electrophoretic mobility data obtained via CE facilitates a better understanding and modeling of physicochemical phenomena.
This method enables the reliable determination of limiting ionic mobilities for various analytes, including aromatic anions.