COSMOSIL HIC

Features

Column Image

Separate based on differences in hydrophobicity
Little loss in enzyme activity and the tertiary structure of proteins

Product Information

Specifications

Packing Material	5HIC
Silica Gel	High Purity Porous Spherical Silica
Average Particle Size	5 µm
Average Pore Size	approx. 300 Å
Specific Surface Area	approx. 150 m²/g
Main Interaction	Hydrophobic Interaction

Applications

A buffer with high salt concentration, usually 1-2 mol/l of (NH4)2SO4, is used as an initial mobile phase for adsorption of samples to a weakly hydrophobic stationary phase. The elution is done with a decreasing salt gradient. The application in lower left shows myoglobin elutes first than BSA under the buffer with high salt concentration, suggesting that myoglobin is less hydrophobic than BSA.

Separation of Protein Standards

Condition
Column Size	4.6 mm I.D. x 50 mm
Mobile Phase	A: 20 mmol/l Phosphate Buffer + 100 mmol/l Na₂SO₄ + 1.5 mol/l (NH₄)₂SO₄ (pH6.7) B: 20 mmol/l Phosphate Buffer + 100 mmol/l Na₂SO₄ (pH6.7) B 0 → 100% /10 min Linear Gradient
Flow Rate	1.0 ml/min
Temperature	30°C
Detection	UV 220 nm
Sample	Myoglobin 1.0 mg/ml β-Lactoglobulin 2.0 mg/ml Hemoglobin 5.0 mg/ml BSA 2.0 mg/ml

Separation of Crude β-Glucosidase

Condition
Column Size	4.6 mm I.D. x 50 mm
Mobile Phase	A: 20 mmol/l Phosphate Buffer + 100 mmol/l Na₂SO₄ + 2 mol/l (NH₄)₂SO₄ (pH6.0) B: 20 mmol/l Phosphate Buffer + 100 mmol/l Na₂SO₄ (pH6.7) B 0 → 100% /10 min Linear Gradient
Flow Rate	1.0 ml/min
Temperature	30°C
Detection	UV 220 nm
Sample	β-Glulosidase 15 µg