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Updated: Mar 23, 2026

Large-scale Top-down Proteomics Using Capillary Zone Electrophoresis Tandem Mass Spectrometry
Published on: October 24, 2018
Three-step stacking by field-enhanced sample injection, sweeping, and micelle to solvent stacking in capillary
Wojciech Grochocki1, Michał J Markuszewski2, Joselito P Quirino3
1Australian Centre for Research on Separation Science (ACROSS), School of Physical Sciences-Chemistry, University of Tasmania, Hobart, Australia; Department of Biopharmaceutics and Pharmacodynamics, Medical University of Gdansk, Gdansk, Poland.
Abstract:
Three-step stacking by field-enhanced sample injection (FESI), sweeping, and micelle to solvent stacking (MSS) in co-EOF capillary zone electrophoresis (CZE) is presented for anionic analytes. Long FESI produced an overloaded stacked zone of analytes (four model penicillins). Sweeping of the FESI zone was by electrokinetic injection of cetyltrimethylammonium bromide (CTAB) micelles. MSS was by short injection of 60% methanol that released the swept analytes from CTAB micelles. The sensitivity enhancement factors were 146-279 and 519-954 for conductivity ratio of 10 and 100, respectively. The SEF enhancement factors (factor=SEF from three-step stacking/SEF from FESI) were 16-32 and 6-10, correspondingly. The LODs were between 6.6-13.2 ng/mL, repeatability (intraday and interday) was %RSD≤5.4%, and linearity was R(2)≥0.998. Application to real sample was investigated using fortified plasma after liquid-liquid extraction.
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