Utilizing Microchip Capillary Electrophoresis Electrospray Ionization for Hydrogen Exchange Mass Spectrometry
Open accessAbstract
Hydrogen exchange (HX) mass spectrometry (MS) of complex mixtures requires a fast, reproducible, and high peak capacity separation prior to MS detection. The current paradigm relies on liquid chromatography (LC) with fast gradients performed at low temperatures to minimize back exchange. Unfortunately, under these conditions, the efficiency of LC is limited due to resistance to mass transfer, reducing the capability to analyze complex samples. Capillary electrophoresis (CE), on the other hand, is not limited by resistance to mass transfer, enabling very rapid separations that are not adversely affected by low temperature. Previously, we have demonstrated an integrated microfluidic device coupling CE with electrospray ionization (ESI) capable of very rapid and high efficiency separations. In this work, we demonstrate the utility of this microchip CE-ESI device for HX MS. High speed CE-ESI of a bovine hemoglobin pepsin digestion was performed in 1 min with a peak capacity of 62 versus a similar LC separation performed in 7 min with peak capacity of 31. A room temperature CE method performed in 1.25 min provided similar deuterium retention as an 8.5 min LC method conducted at 0 °C. Separation of a complex mixture with CE was done with considerably better speed and nearly triple the peak capacity than the equivalent separation by LC. Overall, the results indicate the potential utility of microchip CE-ESI for HX MS.
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Microchip CE-MS analysis of a bovine hemoglobin pepsin digest demonstrated high peak capacity and speed compared to conventional LC separation of the same sample.
“For instance, microchip CE-MS analysis of a bovine hemoglobin pepsin digest has demonstrated high peak capacity and speed when compared to a conventional LC separation of the same sample136.”
From Recent trends of capillary electrophoresis‐mass spectrometry in proteomics research (2019) · cites this paper as [136]
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- Authors
- Black, W. A., Stocks, B. B., Mellors, J. S., Engen, J. R., & Ramsey, J. M.
- Affiliations
- University of North Carolina at Chapel Hill (US)
- Northeastern University (US)
- Corresponding author
- Ramsey, J. Michael
- Venue
- Analytical Chemistry
- Publisher
- American Chemical Society (ACS)
- Published
- 2015-06-02
- Volume
- 87
- Issue
- 12
- Pages
- 6280-6287
- ISSN
- 0003-2700 (ISSN-L)
- 1520-6882
- PMC
- PMC4470751
- OpenAlex
- W281344288
- Cited by
- 35
- References
- 37
- Citations by year
- 2026: 1 · 2025: 2 · 2024: 1 · 2023: 3 · 2022: 3 · 2021: 1 · 2020: 3 · 2019: 3 · 2018: 5 · 2017: 4 · 2016: 6 · 2015: 2
- FWCI
- 2.53
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Locations
- Analytical Chemistry (publishedVersion)
- PubMed (publishedVersion)
- Europe PMC (PubMed Central) (acceptedVersion)
- PubMed Central (acceptedVersion)
- Carolina Digital Repository (University of North Carolina at Chapel Hill) (submittedVersion)
- UNC Libraries
- Funders
- Northeastern University
- Center for Biomedical Microdevices, University of North Carolina, Chapel Hill
- National Institute of General Medical Sciences (R01-GM101135)
- Topics
- Physical Sciences › Chemistry › Spectroscopy › Mass Spectrometry Techniques and Applications
- Physical Sciences › Engineering › Biomedical Engineering › Microfluidic and Capillary Electrophoresis Applications
- Physical Sciences › Chemistry › Spectroscopy › Advanced Proteomics Techniques and Applications
- Keywords
- hydrogen-deuterium exchange mass spectrometry, capillary electrophoresis, electrospray ionization, microfluidics, peak capacity, separation speed, deuterium retention, bovine hemoglobin, pepsin digestion
- MeSH terms
- Animals, Cattle, Chromatography, High Pressure Liquid, Hemoglobins, Mass Spectrometry, Deuterium Exchange Measurement, Electrophoresis, Microchip
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