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Hydrophobic Interaction Chromatography

In HIC, a weakly non-polar stationary phase is used with an aqueous mobile phase containing a high concentration of a chaotropic salt. The technique is mainly applied to the separation of proteins, which are eluted by gradually reducing the salt concentration.
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Hydrophobic Interaction Chromatography

In HIC, a weakly non-polar stationary phase is used with an aqueous mobile phase containing a high concentration of a chaotropic salt. The technique is mainly applied to the separation of proteins, which are eluted by gradually reducing the salt concentration.

As in reversed phase LC, proteins are retained by interaction with alkyl or aryl functional groups on the packing material.

Unlike RPLC, in HIC the density of these functional groups is low and protein molecules are adsorbed on only one or a few sites.

Sorption takes place at high salt concentration, and desorption is accomplished by decreasing the salt concentration or by adding a low percentage of organic solvent.

Although also based on hydrophobic interactions, selectivity in HIC separations is distinctly different from that in reversed phase LC.

Despite the lower peak capacity in HIC compared to RPC, HIC has the advantage that the mobile phase conditions(primarily aqueous) do not usually disrupt higher-order protein structures.

TSKgel HIC Columns

TSKgel HIC packings consist of polymethacrylate base material and a choice of three ligands(butyl, ether and phenyl) with varied hydrophobicities from low to high, respectively.

The polymethacrylate base resin for the ether and phenyl columns is a porous material with a 100 nm pore size and 1,000,000 Da exclusion limit, offering high capacity and high sample loads. The TSKgel Butyl-NPR features a non-porous support in which only the surface of the bead is available for adsorption. Shorter run times and excellent recovery result, making the butyl columns an optimal choice for high throughput or QC analysis.

Figure 1. Comparing Conventional and Nonporous HIC Columns

Greenherbs

TSKgel Phenyl-5PW

Particle Size(mean):

10, 13 and 20 µm

Pore Size(mean):

100 nm

pH Range:

2.0 - 12.0

Functional Group:

Phenyl

P/N

Description

Particle size

Housing material

ID(mm)

Length(cm)

0013063

TSKgel Phenyl-5PW

10 µm

Glass

5

5

0008804

TSKgel Phenyl-5PW

10 µm

Glass

8

7.5

0008808

Guardgel Kit for P/N 0008804

Glass

0018759

TSKgel Phenyl-5PW

10 µm

Stainless Steel

2

7.5

0007573

TSKgel Phenyl-5PW

10 µm

Stainless Steel

7.5

7.5

0007652

Guardgel Kit for P/N 0007573

Stainless Steel

0007656

TSKgel Phenyl-5PW

13 µm

Stainless Steel

21.5

15

0016095

Guardgel Kit for P/N 0007656

Stainless Steel

0020023

TSKgel BioAssist Phenyl

10 µm

PEEK

7.8

5

TSKgel Ether-5PW

Particle Size(mean):

10 µm

Pore Size(mean):

100 nm

pH Range:

2.0 - 12.0

Functional Group:

Ether

P/N

Description

Particle size

Housing material

ID(mm)

Length(cm)

0014013

TSKgel Ether-5PW

10 µm

Glass

5

5

0014014

TSKgel Ether-5PW

10 µm

Glass

8

7.5

0014025

Guardgel Kit for P/N 0014014

Glass

0018760

TSKgel Ether 5PW

10 µm

Stainless Steel

2

7.5

0008641

TSKgel Ether-5PW

10 µm

Stainless Steel

7.5

7.5

0008643

Guardgel Kit for P/N 0008641

Stainless Steel

0008642

TSKgel Ether-5PW

13 µm

Stainless Steel

21.5

15

0016091

Guardgel Kit for P/N 0008642

Stainless Steel

TSKgel Butyl-NPR

Particle Size(mean):

2.5 µm

Pore Size(mean):

non-porous

pH Range:

2.0 - 12.0

Functional Group:

Butyl

P/N

Description

Particle size

Housing material

ID(mm)

Length(cm)

0014947

TSKgel Butyl-NPR

2.5 µm

Stainless Steel

4.6

3.5

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