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| you’re in > Products > Manufacturers > Tosoh > TSK-GEL HILIC COLUMNS |
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TSK-GEL HILIC Columns |
Hydrophilic interaction chromatography (HILIC) is used primarily for the separation of polar and hydrophilic compounds. HILIC has similarities with traditional normal phase chromatography, but the mobile phases for HILIC are similar to those known from reversed phase chromatography (RPC). They include polar organic solvents like methanol or acetonitrile and water.
Compared to RPC the elution order in HILIC mode is inversed for most substances. HILIC is often used to separate hydrophilic compounds such as peptides, carbohydrates and small polar drug candidates or metabolites. Hydrophilic compounds are retained on the polarbonded phase column while nonpolar sample impurities elute unretained in the void volume. In addition it is ideally suited for sensitive LC-MS analysis of water soluble polar compounds because the high organic content in the mobile phase provides rapid evaporation of solvent during electrospray ionization.
The TSK-GEL AMIDE-80 column offers an excellent alternative to amino-bonded stationary phases and consists of 3, 5 or 10 µm silica particles in a stainless steel format. Spherical silica particles are covalently bonded with carbamoyl groups. Based on hydrogen bonds the aqueous content of the mobile phase creates a waterrich layer on the particle surface. This allows for partitioning of polar compounds between the more organic mobile phase and the aqueous layer. The number of polar groups, as well as the conformation and solubility of the sample in the mobile phase determines the elution order. Typical mobile phases consist of acetonitrile buffer mixtures. Samples are eluted from the column by increasing the percentage of the aqueous component.
For years TSKgel Amide-80 columns have been the standard for the analysis of glycans. TSKgel Amide-80 columns packed with 3 µm particles are the newest addition to the TSKgel Amide-80 series. The 3 µm HILIC columns reduce analysis time and improve peak capacity and sensitivity for HPLC and LC-MS analysis. |
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| Applications |
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Small polar molecule drugs:
TSKgel Amide-80 is a valuable tool for the analysis of small, polar
molecule drugs that cannot be retained very well by reversed
phase LC columns.
Figure 1 compares the separation of polar, drug standards with
detection by electrospray ionization mass spectroscopy (ESIMS)
in HILIC mode compared to reversed phase mode. Due to
the high organic content of the eluent HILIC analysis provides
increased detection sensitivity.
Polyalcohols
Polyalcohols are typically separated with a mobile phase of
organic solvent and water as shown in Figure 2 for a 3 μm TSKgel
Amide-80 column compared to a 5 μm column.
Oligosaccharides
The TSKgel Amide-80 can separate oligosaccharides very
rapidly and efficiently. Figure 3 shows a separation of a
ß-cyclodextrin hydrolysate in less than 10 minutes. The labels
indicate the number of base sugars such as glucose in each
oligomer.
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Description |
ID (mm) |
Length (cm) |
Particule Size (μm) |
Number Theoretical Plates |
Flow Rate (mL/min)
|
Max Pressure Drop (kg/cm2)
|
Part # |
| Range |
Max |
| Stainless steel columns |
| Amide-80 |
2.0 |
5.0 |
3 |
≥ 3,500 |
|
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200 |
21864 |
| Amide-80 |
2.0 |
15.0 |
3 |
≥ 13,000 |
|
|
200 |
21865 |
| Amide-80 |
4.6 |
5.0 |
3 |
≥ 6,000 |
|
|
200 |
21866 |
| Amide-80 |
4.6 |
15.0 |
3 |
≥ 18,500 |
|
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200 |
21867 |
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| Amide-80 |
1.0 |
5.0 |
5 |
≥ 300 |
0.03-0.05 |
0.06 |
30 |
20009 |
| Amide-80 |
1.0 |
10.0 |
5 |
≥ 600 |
0.03-0.05 |
0.06 |
60 |
20010 |
| Amide-80 |
1.0 |
15.0 |
5 |
≥ 4,000 |
0.03-0.05 |
0.06 |
90 |
21486 |
| Amide-80 |
1.0 |
25.0 |
5 |
≥ 6,000 |
0.03-0.05 |
0.06 |
120 |
21487 |
| Amide-80 |
2.0 |
5.0 |
5 |
≥ 1,000 |
0.15-0.20 |
0.25 |
40 |
19694 |
| Amide-80 |
2.0 |
10.0 |
5 |
≥ 2,000 |
0.15-0.20 |
0.25 |
80 |
19695 |
| Amide-80 |
2.0 |
15.0 |
5 |
≥ 4,000 |
0.15-0.20 |
0.25 |
100 |
19696 |
| Amide-80 |
2.0 |
25.0 |
5 |
≥ 6,000 |
0.15-0.20 |
0.25 |
150 |
19697 |
| Amide-80 |
4.6 |
5.0 |
5 |
≥ 2,500 |
0.8-1.0 |
1.2 |
50 |
19532 |
| Amide-80 |
4.6 |
10.0 |
5 |
≥ 4,000 |
0.8-1.0 |
1.2 |
50 |
19533 |
| Amide-80 |
4.6 |
25.0 |
5 |
≥ 8,000 |
0.8-1.0 |
1.2 |
150 |
13071 |
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| Amide-80 |
7.8 |
30.0 |
10 |
≥ 5,000 |
1.0-2.0 |
3.0 |
70 |
14559 |
| Amide-80 |
21.5 |
30.0 |
10 |
≥ 8,000 |
4.0-6.0 |
8.0 |
30 |
14460 |
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| Guard column products |
| Amide-80 |
2.0 |
1.0 |
3 |
Cartridge, pk 3 for 2mm ID columns |
21862 |
| Amide-80 |
3.2 |
1.5 |
3 |
Cartridge, pk 3 for 4.6mm ID columns |
21863 |
| Amide-80 |
2.0 |
1.0 |
5 |
Cartridge, pk 3 for 2mm ID columns |
21941 |
| Amide-80 |
4.6 |
1.0 |
5 |
Guard column for 4.6mm ID columns |
19021 |
| Amide-80 |
3.2 |
1.5 |
5 |
Cartridge, pk 3 for 4.6mm ID columns |
19010 |
| Amide-80 |
21.5 |
7.5 |
10 |
Guard column for 21.5mm ID columns |
14461 |
| Cartridge holder For 2.0 mm ID Guard cartridges |
19308 |
| Cartridge holder For 3.2 mm ID Guard cartridges |
19018 |
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NOTE: Tosoh Bioscience offers guard columns and guard
cartridges to protect your analytical column.
Guard cartridges
are usually delivered in packages of three and require the appropriate cartridge holder.
In general cartridges for 4.6 mm ID columns are produced in 3.2 mm ID and 1.5 cm length.
They require the cartridge
holder 19018.
Guard cartridges for 2 mm ID columns are 2 mm
ID x 1 cm L and require holder 19308.
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