Ion Exchange Chromatography

Introduction

Ion-exchange phases separate solutes on the basis of ionic charge. Retention in ion-exchange chromatography is determined by the pH of the eluent, the nature and ionic strength of the buffer and temperature. Column efficiencies are lower than in reversed-phase HPLC. Eluents are normally aqueous but can contain some organic component.

Base Material

Both silica-based and polymer-based ion-exchangers are available. For the former, ionic species are attached to the silica surface, whereas for the latter the ion-exchange groups are distributed throughout the matrix. Silica based materials maintain a mechanical strength and higher efficiency advantage whereas the polymer based materials have greater pH stability.

Classification Application

Ion-exchange is used for the analysis of small ions but its key application area is in the separation of biomolecules such as proteins and nucleic acids. Weak ion-exchangers are used for the analysis of inorganic ions, a technique more specifically termed ion-chromatography.

Ion-Exchange Capacity

The exchange capacity of an ion-exchanger is an important measure of its retentivity (typically measured in milliequivalents per gram material). For any one column the packing density of the phase must also be taken into account. Wide pore materials will typically have lower ion-exchange capacities.

Type

Strength

Nomenclature

Typical Functionality

pH Ionisation Range

Anion

Weak

WAX

Amine

Ionised at specific pH

Anion

Strong

 SAX

Quaternary Ammonium

Ionised over complete pH range

 Cation

 Strong

SCX

Sulphonic Acid

Ionised over complete pH range

 Cation

Weak

WCX

Carboxylic Acid

Ionised at specific pH

Phase

Manufacturer

Base Material

Classification

Particle Size (µm)

Pore Size (Å)

Applications and Features

 Asahipak ES-502N, ES-502C

Shodex

Polymer

WAX, WCX

9

 

Proteins, peptides, oligonucleotides, catecholamines

Shodex IEC QA

 

Polymer

SAX

12

 

Proteins, peptides,

Shodex IEC DEAE, SP, CM

 

Polymer

WAX, SCX, WCX

8

 

Proteins, peptides,

BioBasic AX, SCX

Thermo Scientific

Silica

 

 

 

Proteins, Peptides, Nucleic acids

Capcell Pak UG SCX

Shiseido

 

 

 

 

small molecules

Exsil SAX, SCX

Grace Davison

Polymer

 

 

 

small molecules

Hamilton PRP-X100, PRP-X200

Hamilton

 

 

 

 

Inorganic Ion analysis

Hamilton PRP-X500

Hamilton

 

 

 

 

Proteins, DNA oligomers

Inertsil AX, CX

GL Sciences

 

 

 

 

small molecules

NUCLEOSIL SA, SB 

Macherey-Nagel

 

 

 

 

small molecules

NUCLEOGEN DEAE

Macherey-Nagel

 

 

 

 

Bioanalytical

Partisil SAX, SCX

Whatman

 

 

 

 

small molecules

Partisphere SAX, SCX

Whatman

 

 

 

 

small molecules

PolyCAT A

PolyLC

 

 

 

 

 Aspartic acid functionality

PolySULFOETHYL A

PolyLC

 

 

 

 

Sulfoethylaspartamid

PolyWAX

PolyLC

 

 

 

 

Proteins with isoelectric point <6.0

PL-SAX, PL-SCX

Polymer Labs

Polymer

 

 

 

Protein applications

SynChropak AX, CM

Eprogen

 

 

 

 

Small molecules

SynChropak Q, S

Eprogen

 

 

 

 

Small molecules

TSKgel DEAE-2SW, CM-2SW

Tosoh Bioscience

 

 

 

 

Nucleotides, drug molecules, catecholamines

TSKgel DEAE-3SW, CM-3SW

Tosoh Bioscience

 

 

 

 

Nucleotides, drug molecules, catecholamines

 TSKgel SuperQ-5PW, DEAE-5PW, SP-5PW, CM-5PW

Tosoh Bioscience

 

 

 

 

Enzymes, proteins, DNA, nucleic acids

BioAssist Q

Tosoh Bioscience

 

 

 

 

Plasmids, antibodies and other large proteins

 BioAssist S

Tosoh Bioscience

 

 

 

 

Plasmids, antibodies and other large proteins

 Waters Spherisorb SAX, SCX

Water

Silica

 

 

 

Small molecules

ZirChrom SAX, WAX

ZirChrom Separations

Zirconia

 

 

 

Inorganic and organic anions, biomolecules

ZirChrom SHAX, WCX

ZirChrom Separations

Zirconia

 

 

 

Proteins

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