16
IEC
IEC
TSKgel SW SERIES
The silica-based ion exchange columns are typically used
in the separation of low molecular weight compounds such
as pharmaceuticals, nucleotides or small peptides. Silica-
based particles are used in pore sizes of 125 nanometer
(2SW) and 250 nanometer (3SW) with either diethylami-
noethyl (DEAE) or carboxymethyl (CM) functionality. Bind-
ing capacity for small to medium size proteins on TSKgel
DEAE-3SW is roughly double that of the DEAE-5PW due to
the smaller pore size and larger surface area.
The increased solubility of the silica backbone at pH above
pH 7.5 limits the use of silica based ion exchange columns
to acidic or neutral mobile phases.
Applications of TSKgel SW Ion Exchange Columns
Silica-based Anion Exchange Columns
TSKgel 2SW-type columns provide high performance sepa-
rations of small ionic solutes. High performance analyses
of small anionic species are best performed on small pore
silica-based anion exchangers, such as TSKgel DEAE-2SW.
This is demonstrated in Figure 20. The 250 nanometer pore
size TSKgel DEAE-3SW column is used for separating pep-
tides, low MW proteins and DNA fragments.
Silica-based Cation Exchange Columns
Silica-based cation exchangers are typically used in the
separation of low molecular weight compounds such as
pharmaceuticals, nucleotides, catecholamines, and small
peptides. For example, Figure 21 shows the separation of
nucleosides on TSKgel SP-2SW.
Separation of nucleotides on TSKgel DEAE-2SW
0
24
Minutes
Column:
Sample:
Buffer A:
Buffer B:
Elution:
Flow Rate:
Detection:
TSKgel DEAE-2SW, 4.6mm ID x 25cm
1. AMP, 2. IMP, 3. GMP, 4.ADP, 5. ATP
ACN in 0.1mol/L phosphate, pH 3.0, 20/80
30min linear gradient from buffer A to B
1.0mL/min
UV @ 260nm
1
2
3
4
5
12
ACN in 0.5mol/L phosphate, pH 3.0, 20/80
Separation of nucleotides on TSKgel DEAE-2SW
Column: TSKgel DEAE-2SW, 4.6 mm ID x 25 cm L; Sample: 1. AMP, 2. IMP,
3. GMP, 4.ADP, 5. ATP; Buffer A: ACN in 0.1 mol/L phosphate, pH 3.0, 20/80;
Buffer B: ACN in 0.5 mol/L phosphate, pH 3.0, 20/80; Elution: 30 min linear
gradient from buffer A to B; Flow rate: 1.0 mL/min; Detection: UV @ 260 nm
figure 20
A.
pH 3.5
1
1
2
2
3
3
5
10
Separation of nucleosides by ion-exchange chromatography
on TSKgel SP-2SW
Column:
TSKgel SP-2SW 4.6mm ID x 25cm
Sample:
Nucleoside Standards: 1) Guanosine, 2) Cytidine, 3) Adenosine
Mobile Phase: A) 0.1 mol/L sodium citrate - phosphoric acid buffer, pH 3.5
B) 0.1 mol/L sodium citrate - acetic acid buffer, pH 4.25
Flow Rate:
0.75 mL/min
Temperature: 23°C
Detection:
UV detection @ 260nm
inj.
inj.
0
5
10
0
B.
pH 4.25
Minutes
Minutes
Separation of nucleosides by IEC on TSKgel SP-2SW
Column: TSKgel SP-2SW 4.6 mm ID x 25 cm L
Sample: Nucleoside Standards: 1) Guanosine, 2) Cytidine, 3) A enosine
Mobile Phase: A) 0.1 mol/L sodium citrate - phosphoric acid buffer, pH 3.5
B) 0.1 mol/L sodium citrate - acetic acid buffer, pH 4.25
Flow rate: 0.75 mL/min; Temperature 23 °C; Detection: UV @ 260 nm
figure 21
