Hydrophobic Interaction Chromatography (HIC) Media

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Introduction


HIC is a versatile technique and can show high selectivity to individual molecules according to their exposed hydrophobic zones. It is particularly useful for intermediate and final-stage purifications.

Butyl SepFast™, Phenyl SepFast™, and Octyl SepFast™ are hydrophobic interaction chromatography (HIC) adsorbents with carbon chains of C4, C6 (benzene ring), and C8, respectively. They are specially designed for the purification of biological molecules based on their hydrophobicity profiles.

HIC media requires much milder purification conditions than reversed phase chromatography (RPC) media. Hence biological activity can be maintained in HIC separations unlike RPC operations.

HIC media normally binds at moderate to high salt concentrations. It is therefore logical to place a HIC step after an IEX step where molecules are usually eluted at high salt conditions.

SepFast™ HIC media offers a broad range of choices from C4 to C8. Generally speaking, the longer the carbon chain, the higher the surface hydrophobicity. 

Biotoolomics HIC media gives you:

  • High sample loading capacity
  • High separation power
  • Available at all pack sizes from analytical up to full manufacturing scale.
  • Chemically stable Agarose base matrix.
  • Can be produced in high and low ligand densities, alternate crosslinking levels and different bead sizes to optimise the media to the separation.
  • All versions can be scaled up and produced for GMP use
  • More choices than other suppliers.

The feature and selection guide is listed as follows:

4HF serial
(50m - 150m)
6HF serial
(50m - 150m)
Large Beads
(150m - 350m)
HighRes
(25m - 50m)
The above HIC media is designed to purify most medium to large proteins The above HIC media is designed to purify peptides or smaller proteins The above HIC media is designed to purify proteins from crude or viscous samples The above HIC media is designed to purify proteins
that require high resolution

Characteristics of SepFast HIC Media

 
Ligand Butyl Phenyl Octyl
Matrix Highly cross-linked agarose; 4% agarose for SepFast 4HF, 6% agarose for SepFast 6HF
Functional
group
-(CH2)3CH3 -C6H6 -(CH2)7CH3
Ligand
density
Around 40 mol/ml Around 25 mol/ml;
Around 40 mol/ml (high sub)
Around 4 mol/ml
Binding
capacity
>20 mg lyzosyme / ml >20 mg BSA / ml >20 mg BSA / ml
Particle
size
25 - 50 m (HighRes), 50 - 150 m (4HF and 6HF), 150 - 350 m (Large Beads)
Pressure
-flow
property*
>100 cm/h for HighRes; >300 cm/h for 4HF serials; >500 cm/h for 6HF serials; >1500 cm/h for Large Beads
Operational
pressure
Up to 3 bar
pH
stability
2-14 (short term) and 3-12 (long term)
Working
temperature
4oC – 30oC
Chemical
stability
All commonly used buffers; 1 M acetic acid, 1 M NaOH, 6M guanidine hydrochloride, 8 M urea, 30% acetonitrile, 30% isopropanol, 70% ethanol, 2 M (NH4)2SO4
Storage 20% ethanol

*Measured in a 32 mm ID column at a bed height of 15 cm.

Ordering information

 
HIC Selection Kit
Description Quantity Code no. Price GBP
HIC SepFast Selection Kit 7 x 1 ml column packed with Butyl 4HF, Butyl 6HF, Phenyl 4HF, Phenyl 6HF, Phenyl 6HF high sub, Octyl 4HF and Octyl 6HF, respectively 480000-7x1ML 169.00
SepFast Phenyl Products SepFast Octyl Products
Butyl Products
Description Quantity Code no. Price GBP
Butyl SepFast 4HF 25 ml 480101-25ML 94.92
  100 ml 480101-100ML 250.01
  1 Litre 480101-1L POR
Butyl SepFast 4HF prepacked column 5 x 1 ml 480101-5x1ML 129.00
  5 x 5 ml 480101-5x5ML 268.00
Butyl SepFast 6HF 25 ml 480301-25ML 94.92
  100 ml 480301-100ML 250.01
  1 Litre 480301-1L POR
Butyl SepFast 6HF prepacked column 5 x 1 ml 480301-5x1ML 129.00
  5 x 5 ml 480301-5x5ML 268.00
Butyl SepFast Large Beads 25 ml 480201-25ML 94.92
  100 ml 480201-100ML 250.01
  1 Litre 480201-1L POR
SepFast Butyl Products SepFast Octyl Products
Phenyl Products
Description Quantity Code no. Price GBP
Phenyl SepFast 4HF 25 ml 481001-25ML 94.92
  100 ml 481001-100ML 250.01
  1 Litre 481001-1L POR
Phenyl SepFast 4HF prepacked column 5 x 1 ml 481001-5x1ML 129.00
  5 x 5 ml 481001-5x5ML 268.00
Phenyl SepFast 6HF 25 ml 481201-25ML 94.92
  100 ml 481201-100ML 250.01
  1 Litre 481201-1L POR
Phenyl SepFast 6HF prepacked column 5 x 1 ml 481201-5x1ML 129.00
  5 x 5 ml 481201-5x5ML 268.00
Phenyl SepFast 6HF High sub 25 ml 481202-25ML 94.92
  100 ml 481202-100ML 250.01
  1 Litre 481202-1L POR
Phenyl SepFast 6HF High sub prepacked column 5 x 1 ml 481202-5x1ML 129.00
  5 x 5 ml 481202-5x5ML 268.00
Phenyl SepFast 6HF HighRes 25 ml 481101-25ML 115.00
  100 ml 481101-100ML 300.00
  1 Litre 481101-1L POR
Phenyl SepFast 6HF HighRes prepacked column 5 x 1 ml 481101-5x1ML 154.00
  5 x 5 ml 481101-5x5ML 294.00
Phenyl SepFast Large Beads 25 ml 481301-25ML 94.92
  100 ml 481301-100ML 250.01
  1 Litre 481301-1L POR
SepFast Butyl Products SepFast Phenyl Products
Octyl Products
Description Quantity Code no. Price GBP
Octyl SepFast 4HF 25 ml 481601-25ML 94.92
  100 ml 481601-100ML 250.01
  1 Litre 481601-1L POR
Octyl SepFast 4HF prepacked column 5 x 1 ml 481601-5x1ML 129.00
  5 x 5 ml 481601-5x5ML 268.00
Octyl SepFast 6HF 25 ml 481801-25ML 94.92
  100 ml 481801-100ML 250.01
  1 Litre 481801-1L POR
Octyl SepFast 6HF prepacked column 5 x 1 ml 481801-5x1ML 129.00
  5 x 5 ml 481801-5x5ML 268.00
Octyl SepFast Large Beads 25 ml 481701-25ML 94.92
  100 ml 481701-100ML 250.01
  1 Litre 481701-1L POR


>>Click here for SepFast Butyl Products

>>Click here for SepFast Phenyl Products

>>Click here for SepFast Octyl Products

Note: HIC media with lower or higher ligand density can be custom-made to increase recovery or binding capacity. Please contact us for more information.

How to order

Data Sheet

Data sheet