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Ion-Exchange Chromatography01:09

Ion-Exchange Chromatography

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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...
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Ion Exchange01:17

Ion Exchange

1.5K
Ion exchange chromatography separates charged molecules from a solution by reversibly exchanging them with mobile, or 'active', ions associated with the oppositely charged stationary phase. This method can be used to separate ions, soften and deionize water, and purify solutions. The polymers comprising the ion-exchange column are high-molecular-weight and chemically stable polymers, crosslinked to be porous and essentially insoluble. They are also functionalized with either acidic or...
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High-Performance Liquid Chromatography: Elution Process01:05

High-Performance Liquid Chromatography: Elution Process

1.8K
In High-Performance Liquid Chromatography (HPLC), the elution process is critical to the separation of analytes and the quality of chromatographic results. Elution describes how compounds move through the column and separate based on their interactions with the mobile and stationary phases. This process determines the resolution, peak shape, and retention times in the chromatogram, which are essential for identifying and quantifying components in complex mixtures. Understanding the elution...
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Size-Exclusion Chromatography01:08

Size-Exclusion Chromatography

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In size-exclusion chromatography (SEC), also known as molecular-exclusion or gel-permeation chromatography, molecules are separated based on their sizes. This technique is important for separating large molecules such as polymers and biomolecules. The two classes of micron-sized stationary phases encountered in SEC are silica particles and cross-linked polymer resin beads. Both materials are porous, but their pore sizes vary significantly.
Silica particles offer advantages such as rigidity,...
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Capillary Electrophoresis: Applications01:30

Capillary Electrophoresis: Applications

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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,...
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High-Performance Liquid Chromatography: Introduction01:11

High-Performance Liquid Chromatography: Introduction

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High-performance liquid chromatography(HPLC), formerly referred to as High-pressure liquid chromatography, is a powerful technique used to separate, identify, and quantify components in complex mixtures. The term "high pressure" refers to using high pressure to push the liquid mobile phase through the tightly packed columns.
In HPLC, two phases play a critical role in the separation process:
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Related Experiment Video

Updated: Mar 12, 2026

Ion Exchange Chromatography IEX Coupled to Multi-angle Light Scattering MALS for Protein Separation and Characterization
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Ion Exchange Chromatography IEX Coupled to Multi-angle Light Scattering MALS for Protein Separation and Characterization

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Heparin Isomeric Oligosaccharide Separation Using Volatile Salt Strong Anion Exchange Chromatography.

Rebecca L Miller1,2, Scott E Guimond1, Maitreyi Shivkumar1

  • 1Centre for Glycobiology, Department of Biochemistry, Institute of Integrative Biology, University of Liverpool , Crown Street, Liverpool L69 7ZB, United Kingdom.

Analytical Chemistry
|November 2, 2016
PubMed
Summary
This summary is machine-generated.

Purifying complex heparin and heparan sulfate saccharides is challenging. A new volatile salt buffer method (VSCTA-SAX) improves separation and purity, enabling better structure-activity studies.

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Area of Science:

  • Glycoscience
  • Analytical Chemistry
  • Biochemistry

Background:

  • Heparin and heparan sulfate saccharide purification is complex due to numerous isomeric structures.
  • Current purification methods (SAX chromatography) face challenges like sample loss during desalting and incompatibility with mass spectrometry.

Purpose of the Study:

  • To develop an improved purification method for heparin and heparan sulfate saccharides.
  • To enhance resolution and purity of saccharide isomers for structure-activity relationship studies.

Main Methods:

  • Utilized cetyltrimethylammonium-strong anion exchange (CTA-SAX) chromatography with a volatile salt (VS) buffer, specifically ammonium bicarbonate.
  • Employed ion mobility mass spectrometry to analyze structural conformations of isomeric saccharides.

Main Results:

  • The volatile salt CTA-SAX (VSCTA-SAX) method demonstrated improved resolution of isomeric saccharide structures compared to standard SAX methods.
  • This method provides an orthogonal separation technique, yielding higher purity saccharides.
  • The use of ammonium bicarbonate allows for sample evaporation, preventing loss and facilitating subsequent mass spectrometry analysis.

Conclusions:

  • VSCTA-SAX is a powerful new tool for separating challenging heparin/heparan sulfate saccharide mixtures.
  • This technique overcomes limitations of previous methods, reducing sample loss and improving compatibility with mass spectrometry.
  • The enhanced separation capabilities will significantly advance structure-activity studies of these complex biomolecules.