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Hydrophobic Interaction Chromatography (HIC)
Hydrophobic interaction chromatography (HIC) is a special method used for the separation of biomolecules by utilising their hydrophobic properties. This method plays a central role in the purification and analysis of proteins, peptides and other macromolecules, especially when the preservation of the native structure and function of the molecules is important.
The separation is based on the interaction of hydrophobic regions of the proteins with a weakly hydrophobic stationary phase. The analytes are therefore separated according to their degree of surface polarity. Proteins with high surface hydrophobicity are more strongly retained by the stationary phase than those whose hydrophobic regions are found more in the interior of the protein. These interactions between protein and stationary phase are also greatly increased by the addition of salts to a buffered mobile phase, as the solubility of the proteins is reduced by the presence of the salts and thus the hydrophobic interactions of the protein with the stationary phase increase. The proteins are eluted from the column by a descending salt gradient. The mobile phase is purely aqueous and free of organic solvents, as these can lead to denaturation of the proteins to be analysed.
Here you will find an overview of manufacturers and HILC-HPLC columns. We will be happy to assist you in selecting the right column for your analytical problem!
Technical Data
Manufacturer and columns of hydrophobic interaction chromatography
Shodex™
| Name | Functional Group | Particle Size | Pore Size | Column Size |
| HIC PH-814 | Phenyl | 10 µm | 2000 Å | 8.0 x 75 mm |
Base material: Polyhydroxymethacrylate
Tosoh Bioscience™
| Name | Functional Group | Particle Size | Pore Size | Column Size |
| TSKgel Ether 5-PW | Polyether | 10 µm | 1000 Å | 5.0 x 50 mm[1] |
| TSKgel Phenyl 5-PW | Phenyl | 10 µm | 1000 Å | 5.0 x 50 mm[1] |
| TSKgel BioAssist Phenyl | Phenyl | 10 µm | 1000 Å | 7.8 x 50 mm[4] |
| TSKgel Butyl-NPR | Butyl | 2.5 µm | Non-porous | 4.6 x 35 mm[2] |
Base material: polymethacrylates; [1] glass columns; [2] stainless steel columns; [3] particle size 13 µm; [4] PEEK column
Thermo Scientific™
| Name | Functional Group | Particle Size | Pore Size | Column Size |
| MAbPac HIC-10[1] | Proprietary polyamide | 5 µm | 1000 Å | 4.6 x 100 mm |
| MAbPac HIC-20[1] | Proprietary alkylamide | 5 µm | 1000 Å | 4.6 x 100 mm |
| MAbPac HIC-Butyl[2] | Butyl | 5 µm | Non-porous | 4.6 x 100 mm |
| ProPac HIC-10[1] | Proprietary alkylamide | 5 µm | 300 Å | 7.8 x 75 mm |
[ 1] Base material: Silica gel; [2] Base material: Hydrophilic polymer
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