Related Experiment Video
Updated: May 28, 2026

Capillary Electrophoresis Separation of Monoclonal Antibody Isoforms Using a Neutral Capillary
Published on: January 16, 2017
[Comparison of different buffer systems for separation of 15 nucleosides by capillary electrophoresis]
Qian Shi1, Junhui Chen, Xin Li
1Qingdao Key Lab of Analytical Technology and Standardization of Chinese Medicines, The First Institute of Oceanography, State Oceanic Administration, Qingdao 266061, China.
Abstract:
The most suitable background electrolytes (BGEs) for simultaneous separation of 15 nucleosides by different modes of capillary electrophoresis (CE) were obtained. Various modes of CE were performed including capillary zone electrophoresis (CZE), capillary electrophoresis-electrospray ionization-time of flight mass spectrometry (CE-ESI-TOF/MS) and micellar electrokinetic chromatography (MEKC). The electrolyte buffers using sodium tetraborate decahydrate, disodium hydrogen phosphate, sodium acetate, sodium bicarbonate, ammonium acetate or 1, 2-diamino-ethane (DEA) were tested, and the best of them were systematically optimized. In CZE mode, the nucleosides could not be separated completely with sodium tetraborate decahydrate or disodium hydrogen phosphate as BGEs, demonstrating the limited applicability of the two buffer systems for complex samples. However, with 300 mmol/L DEA (containing 2% acetone) as BGE, 15 nucleosides could be separated with good resolution and peak shape, which proved that the DEA buffer was most suitable in CZE. The best buffer system in MEKC mode was 25 mmol/L disodium hydrogen phosphate with 70 mmol/L sodium dodecyl sulfate (SDS), and it was successfully applied for the separation of the nucleosides in Chinese Anthopleura lanthogrammica Berkly. The optimum buffer system for CE-ESI-TOF/MS analysis was 20 mmol/L ammonium acetate (pH 10.0). In the positive ion mode, the MS signals of each compound were better than those in the literature using DEA as BGE. The results of this study demonstrated the applicability of different buffer systems for the simultaneous separation of 15 nucleosides, and were helpful for the development of CE method in complex sample separation.
Related Concept Videos
Capillary Electrophoresis: Applications
Capillary zone electrophoresis (CZE) separates ionic components based on their electrophoretic mobility. It has been used to separate proteins, amino acids,...
Capillary Electrophoresis: Instrumentation
Electrophoresis: Overview
There...
Ion-Exchange Chromatography
SDS-PAGE
A variation of gel electrophoresis, termed polyacrylamide gel electrophoresis (PAGE), is commonly used for separating proteins according to their molecular size by passing them through a polyacrylamide gel. Because of the varying charges associated with amino acid side chains, PAGE can be used to separate intact proteins...
Two-dimensional Gel Electrophoresis
The first dimension separation uses the isoelectric focusing or IEF technique performed on immobilized pH gradient (IPG) strips that separate proteins according to their isoelectric points.
Biological samples, such as cells...
