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Size Exclusion LC

Size Exclusion Chromatography (SEC)

Introduction

SEC is about separation of components according to their molecular size in solution, larger molecules eluting first. Separation is achieved by the differential exclusion or inclusion of components within the packing material particles. In addition to the separation of discrete components, the technique is used for characterizing the molecular weight distribution of polymers.

Gel Permeation Chromatography (GPC)

Modes of Operation

Gel permeation chromatography (GPC) refers to the separation of organic soluble polymers using an organic solvent as the eluent.

Base Material

Silica based SEC materials generally exhibit higher resolving power than polymer based materials. However, polymer based materials show greater stability for use with high pH eluents. Polymeric packing materials are generally available in larger particle sizes, which may be more practical for large scale preparative separations.

Applications

SEC analyses do not normally result in the denaturation of samples, making the technique a suitable choice for biological samples where activity must be retained. A wide range of biomolecules and organic polymers are separated by SEC. For samples of wide molecular weight distribution, it can be useful to use a mixed pore size phase or to couple columns of one or more pore sizes in series.

Gel filtration chromatography (GFC)

Gel filtration chromatography (GFC) refers to the separation of water soluble polymers in aqueous eluents. SEC separations exhibit lower resolving power and capacity compared with adsorptive HPLC techniques.

 

SEC Phases

Phase /Series

Manufacturer

Base Material / Bonding

Mode

Pore Size (A)

Typical Applications

GPC Peptide

Eprogen

Glycerol Bonded Silica

GFC

50

Small peptides

GPC100,300,500,1000,4000

GFC

100, 300, 500, 1000, 4000

Proteins, carbohydrates, nucleic acids, water soluble polymers

GPC Linear

GFC

100 - 1000

Organic polymers, denaturated polymers

CAT SEC

Sillica with Polymerised Polyamine

GFC

100, 300, 1000,

Cationic Polymers

MCI GEL CQP

Mitsubishi Chemical Corp

Polyhydroxymethacrylate

GFC

120, 200, 600,

Proteins, peptides, enzymes, and other biomolecules

PLgel

Agilent Technologies

Polystyrene Divinylbenzene

GPC

50, 100, 500, 1000, 10 000, 100’000 Mixed

Oils, oligomers, high MW synthetic polymers, starches, gum

PL Aquagel-OH,30,40,50,60,Mixed

Polystyrene Divinylbenzene with Polyhydroxyfunctionality

GFC

-

Surfactants, polysaccarides, polyacrylamides, starches, gum

Polyhydroxyethyl A

PolyLC

Silica with hydroxymethylaspartamide coating

GFC

60, 100, 200, 300, 500, 1000, 1500

Peptides, proteins, carbohydrates, small molecules,

Asahipak GF

Showa Denko

Polyvinyl alkohol

GFC / GPC

400, 2000, 10000

Hydrophilic and hydrophobic compounds

Shodex GPC

Polystyrene Divinylbenzene

GPC

various

Polymers, plastics

Shodex OHpak SB

Polyhydroxymethacrylate

GFC

various

Water soluble samples

Shodex Protein KW

Silica

GFC

400, 1000, 1500

Proteins

Acclaim SEC

Thermo Scientific

Hydrophilic Polymethacrylate

GFC

300, 1000

Water soluble polymers

BIOBasic SEC

Silica

GFC

60, 120, 300, 1000,

Peptides and Proteins

MAbPac SEC1

Silica

GFC

300

Monoclonal antibodies and aggregates

TKSgel SW

Tosoh Bioscience

Silica

GFC

125, 250, 450

Proteins, antibodies, enzymes, nucleic acids

TKSgel PW

Polyhymethacrylate

GFC

<100, 125, <200, 200, 500, 1000, >1000

 Water soluble organic polymers, polysaccarides, DNA

TKSgel H

Polystyrene Divinylbenzene

GPC

-

Oligomers, polymers and polymer additives

TKSgel Alpha and Super AW

Hydrophilic Polyvinyl

GFC /GPC

-

Organic and water soluble polymers

Zorbax GF series

Agilent Technologies

Zirconia clad Silica

GFC

150, 300

Proteins, Peptides

 

 

 

 

 

 

 

 

 

 

 

 

 

 

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