78
RPC
Reversed phase chromatography (RPC) columns can be applied to the
analysis of a wide variety of compounds, ranging from neutral polar
and non-polar solutes to acidic, basic and amphoteric compounds.
RPC is also and efficient technique for the analysis of derivatized amino
acids, peptides and proteins, although protein structure is not always
maintained due to the high concentration of organic solvent required
for elution. Tosoh Bioscience offers 18 distinct RPC column types which
are based on either silica or polymer particles (
Table I
).
The silica-based TSKgel RPC product line consists of ten stationary
phases designed for the analysis of low molar mass compounds,
including active pharmaceutical ingredients (API), forensic compounds,
derivatized amino acids, carbohydrates, steroids, lipids, and fatty acids,
as well as two stationary phases with larger pore size designed for
protein analysis.
TSKgel silica packings consist of spherical particles with uniform pore
sizes of 8, 10, 12, 14, 25, or 30 nm bonded with a monomeric or polymeric
layer of octadecyl, octyl, cyano, trimethylsilyl, or phenyl groups. Several
of the stationary phases are subsequently endcapped by derivatization
with trimethylsilyl groups by a proprietary method that deactivates
residual silanol groups.
Polymethacrylate-based reversed phase columns are available in a
range of pore and particle sizes. Although often not as efficient as
and less robust than silica-based RPC columns, key advantages of
polymer-based columns are their pH stability from pH 2 to 12, which
allows many basic compounds to be analyzed in their uncharged form,
thus reducing secondary adsorption and improving peak shape and
improving recovery for peptides and proteins due to reduced secondary
interactions.
TSKgel REVERSRED PHASE CHROMATOGRAPHY COLUMNS
Silica-based columns
Polymer-based columns
High purity type B silica
Hydrophilic backbone to improve recovery and reduce secondary
interactions
High efficiencies
pH stable from 1 to 12
Excellent recoveries
Compatability with organic solvents eliminates swelling
Low bleed for MS
An excellent choice for analysis of small molecules and peptides
An excellent choice for large MW biomolecules and for analyzing
small MM compounds at high pH
TSKgel Protein C
4
-300
TSKgel Octadecyl-2PW
High efficiency & throughput TSKgel ODS-100V and 100Z
TSKgel Octadecyl-4PW
Monomeric bonded silica
TSKgel Phenyl-5PW RP
Specialty silica columns
TSKgel Octadecyl-NPR (nonporous)
TABLE I
Silica and polymer based TSKgel RPC columns
