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Related Concept Videos

Capillary Electrophoresis: Instrumentation01:20

Capillary Electrophoresis: Instrumentation

Capillary electrophoresis instrumentation typically consists of several key components. A high-voltage power supply generates the electric field necessary for the separation by connecting to an anode (the positively charged electrode) and a cathode (the negatively charged electrode) located in buffer reservoirs at each end of the capillary tube. The system includes a sample vial, a fused silica capillary tube coated with polyimide for mechanical strength through which the sample components...
Capillary Electrophoresis: Applications01:30

Capillary Electrophoresis: Applications

Capillary electrophoretic separations offer various modes, each with unique applications. These modes include capillary zone electrophoresis, capillary gel electrophoresis, capillary array electrophoresis, capillary isoelectric focusing, capillary isotachophoresis, micellar electrokinetic chromatography, and capillary electrochromatography.
Capillary zone electrophoresis (CZE) separates ionic components based on their electrophoretic mobility. It has been used to separate proteins, amino acids,...
Electrophoresis: Overview01:20

Electrophoresis: Overview

Electrophoresis is a powerful analytical separation technique that relies on the differential migration of charged species when subjected to an electric field. The core strength of electrophoresis lies in its ability to separate high-molecular-weight species in complex mixtures. It has found widespread use in biochemistry, molecular biology, and analytical chemistry, allowing the separation of compounds like amino acids, nucleotides, carbohydrates, and proteins with excellent resolution.
There...
Ion-Exchange Chromatography01:09

Ion-Exchange Chromatography

Ion-exchange chromatography, or IEC, is a technique for separating ions based on their affinity for the stationary phase. The stationary phase is a cross-linked polymer resin with covalently attached ionic functional groups. The functional groups can be either positively charged (cation exchangers) or negatively charged (anion exchangers). A cation exchanger consists of a polymeric anion and active cations, while an anion exchanger is a polymeric cation with active anions. The choice of...
Two-dimensional Gel Electrophoresis01:22

Two-dimensional Gel Electrophoresis

Two-dimensional gel electrophoresis is a high-resolution protein separation method first introduced by O' Farrell and Klose in 1975. This method involves protein separation by two dimensions, mass and charge, making it more accurate than one-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...
Electrospray Ionization (ESI) Mass Spectrometry01:12

Electrospray Ionization (ESI) Mass Spectrometry

Higher molecular weight biomolecules are nonvolatile compounds that may decompose before ionizing or vaporizing during mass analysis with conventional electron impact ionization methods. Accordingly, electrospray ionization (ESI) is the favored method for vaporizing and ionizing biomolecules as it circumvents rapid fragmentation and enables the recording of mass signals for the entire biomolecule.
ESI utilizes electrical energy to transfer ions from the liquid phase of the sample into the...

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Optimization of capillary zone electrophoresis/electrospray ionization parameters for the mass spectrometry and tandem mass spectrometry analysis of peptides.

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Updated: Jun 1, 2026

Large-scale Top-down Proteomics Using Capillary Zone Electrophoresis Tandem Mass Spectrometry
10:05

Large-scale Top-down Proteomics Using Capillary Zone Electrophoresis Tandem Mass Spectrometry

Published on: October 24, 2018

Ultrahigh-voltage capillary zone electrophoresis.

K M Hutterer1, J W Jorgenson

  • 1Department of Chemistry, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 27599-3290.

Analytical Chemistry
|June 14, 2011
PubMed
Summary
This summary is machine-generated.

Researchers developed an ultrahigh-voltage capillary electrophoresis system, achieving millions of theoretical plates for peptide separation by increasing applied voltage to 120 kV. This advancement significantly enhances separation power in capillary electrophoresis.

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Highly Sensitive and Quantitative Detection of Proteins and Their Isoforms by Capillary Isoelectric Focusing Method
07:58

Highly Sensitive and Quantitative Detection of Proteins and Their Isoforms by Capillary Isoelectric Focusing Method

Published on: September 19, 2018

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Large-scale Top-down Proteomics Using Capillary Zone Electrophoresis Tandem Mass Spectrometry
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Published on: October 24, 2018

Highly Sensitive and Quantitative Detection of Proteins and Their Isoforms by Capillary Isoelectric Focusing Method
07:58

Highly Sensitive and Quantitative Detection of Proteins and Their Isoforms by Capillary Isoelectric Focusing Method

Published on: September 19, 2018

Area of Science:

  • Analytical Chemistry
  • Separation Science
  • Biochemistry

Background:

  • Capillary electrophoresis (CE) is a powerful separation technique.
  • Enhancing separation power in CE is crucial for complex sample analysis.
  • Limitations exist in achievable separation efficiency with conventional voltage limits.

Purpose of the Study:

  • To demonstrate the feasibility of extending applied potential in capillary electrophoresis.
  • To increase the separation power of capillary electrophoresis systems.
  • To investigate the impact of ultrahigh voltages on CE performance.

Main Methods:

  • Modification of a commercial 30-kV power supply to achieve up to 120 kV.
  • Development of a unique electrical shielding system to prevent discharge.
  • Electrophoretic separation of peptide standards and protein digests.

Main Results:

  • Achieved theoretical plate counts ranging from 2.7 to 6.1 million for peptides at 120 kV.
  • Demonstrated a linear increase in theoretical plates with applied voltage.
  • Observed resolution increasing with the square root of applied voltage, consistent with theory.

Conclusions:

  • Ultrahigh-voltage capillary electrophoresis significantly enhances separation efficiency.
  • The developed system overcomes breakdown issues associated with high voltages.
  • This approach offers a pathway to dramatically improve CE separation power for complex mixtures.