SEC
8
INTRODUCTION TO TSKgel SIZE EXCLUSION COLUMNS
Size Exclusion Chromatography (SEC) is the dominant mode of
separation for polymers. SEC is the general name for the
chromatographic mode in which components of a mixture are
separated according to their molecular size, based on the flow of
the sample through a column packed with porous particles. Large
sample molecules cannot or can only partially penetrate the pores,
whereas smaller molecules can access all or a larger number of
pores. In SEC, large molecules elute from the column first followed
by smaller molecules, and the smallest molecules that can access all
the pores elute last from the column. Size exclusion chromatography
is the only mode of chromatography that does not involve interaction
with a stationary phase by means of adsorption or partitioning of the
solutes.
The terms SEC, GFC (gel filtration chromatography) and GPC (gel
permeation chromatography) all refer to the same chromatographic
technique. In GFC, an aqueous mobile phase is used, while an organic
mobile phase is employed in GPC. The general term SEC covers both
uses. Available TSKgel products are classified by application area
and particle composition.
GEL FILTRATION CHROMATOGRAPHY (GFC)
The principal feature of GFC is its gentle non-interaction with the
sample, enabling retention of enzymatic activity while separating
multimers that are not easily distinguished by other chromatographic
methods. SEC has limited peak capacity, however, requiring that
the molar mass of the biomolecules differ by at least twofold. GFC
is popular among biochemists for the isolation of proteins, for the
removal of aggregates, to desalt a protein sample, to separate nucleic
acid fractions, or to characterize water soluble polymers used in food
products, paints, pharmaceutical preparations, etc.
TSKgel columns for GFC analysis consist of the TSKgel SW and PW
series column lines. The main criterion in choosing between these
TSKgel columns is the molar mass of the sample and its solubility. The
fact that the TSKgel SW columns are based on silica and the TSKgel
PW columns are derived from a hydrophilic polymer network has less
impact on the separation than the particle and pore size differences
between the column lines. While a TSKgel SW column is typically
the first column to try for biopolymers, TSKgel PW columns have
demonstrated good results for smaller peptides (<1,000 Da), protein
aggregates, DNA fragments, and viruses.
Application area:
Proteins and other biopolymers
Base material: silica
• SW
• SW
XL
• SuperSW/SuperSW mAb
• UltraSW
Due to higher resolving power, the TSKgel SW series columns are
ideal for proteins and nucleic acids using an aqueous buffer as mobile
phase. The TSKgel SWmAb columns within the TSKgel SW series are
designed specifically for the analysis of monoclonal antibodies.
Application area:
Water soluble polymers
Base material: polymethacrylate
• PW
• SuperMultiporePW
• SuperOligoPW
• PW
XL
• PW
XL
-CP
TSKgel PW series columns are commonly used for the separation
of synthetic polymers, oligosaccharides, nucleic acids and small
viruses using aqueous buffer or salt solutions as mobile phase. The
TSKgel SuperMultiporePW semi-micro SEC columns provide near
linear calibration curves and are ideally suited to analyze the MW
distribution of water soluble polymers with a wide range of molecular
weights. The SuperOligoPW semi-micro column featuring a small
particle size has been designed for fast analysis of oligosaccharides
and other oligomers. The PW
XL
-cP columns are developed to facilitate
SEC separation of cationic polymer under low salt conditions.
FEATURES
BENEFITS
Rigid hydrophilic and hydrophobic packings
Minimal swelling and excellent physical strength
Low adsorption resulting in high mass recovery
Four series of SEC columns with diferent ranges of solvent
compatibility
Suitable for both types of size exclusion, aqueous (GFC) and
non-aqueous (GPC)
Easy scale up
Analytical and preparative pre-packed SEC column
