COSMOSIL Protein-R

Features

Column Image

Excellent separation
High recovery rate
Outstanding stability at low pH

Product Information

Specifications

Packing Material	Protein-R
USP Code	L1
Silica Gel	High Purity Porous Spherical Silica
Average Particle Size	5 µm
Average Pore Size	approx. 300Å
Specific Surface Area	approx. 150 m²/g
Stationary Phase	Octadecyl Group
Bonding Type	Polymeric
Main Interactions	Hydrophobic Interaction
End Capping Treatment	Near-perfect Treatment
pH Range	1.5-7.5*
Feature	- High recovery rate - Acid-resistance

* Optimal pH range of silica-based columns is between 2 and 7.5. Extreme pH may significantly decrease column lifetime.

Comparison of Separation

Protein-R shows sharper peaks for proteins than conventional C18 wide pore columns.

Protein-R

Comparison of separation

5C₁₈-300

Comparison of separation

Condition
Column Size	4.6 mm I.D. x 150 mm	Sample	Ribonuclease A 1.0 µg Cytochrome C 1.0 µg Lysozyme 1.0 µg BSA 1.0 µg Myoglobin 1.0 µg Ovalbumin 1.0 µg
Mobile Phase	A : 0.05%TFA - 20% Acetonitrile
	B : 0.05%TFA - 60% Acetonitrile
	B conc. 0 → 35% 60 min Linear Gradient
Flow Rate	1.0 ml/min
Temperature	30°C
Detection	UV 220 nm

Recovery Rate

The figure below shows recovery rates for proteins using different columns. Protein-R shows a higher recovery rate than C₄-300 and a much higher recovery rate than C₁₈-300.

recoveryrate

Durability

The figure below shows durability against acidic mobile phase of various columns. Protein-R shows a higher acid durability than C₄-300.

durability

Protein Separation Wide Pore Columns

Reversed Phase Chromatography	Protein-R C₁₈-AR-300･C₈-AR-300･C₄-AR-300･Ph-AR-300
Gel Filtration Chromatography	Diol-120-II ･Diol-300-II
Hydrophobic Interaction Chromatography	HIC

Applications

Peptide Separation

For low-MW peptides, 5C₁₈-AR-II (pore size: 120 Å) performs better; however, for high-MW peptides, Protein-R yields better separation.

peptide separation chromatogram

Condition
Column Size	4.6 mm I.D. x 150 mm	Sample	Poly-L-Glutamic Acid Sodium Salt, MW 1500-5000 (200µg)
Mobile Phase	A: 0.05% TFA-H₂O
	B: 0.05% TFA-MeCN
	B conc. 0 → 35% / 60 min linear gradient
Flow Rate	1.0 ml/min
Temperature	30°C
Detection	UV 220 nm

Usage information

Column	5 µm Protein-R
I.D. (mm)	2.0	4.6	10.0	20.0
Washing method	Washing method 1. Remove buffer, salts and/or acid from column: wash for 10-15 min. using the mobile phase last used, without buffer, salts or acid. 2. (If you used a salt/acid, always do step 1 first!) Remove adsorbed compounds and fix unstable baselines by washing with 50-70% acetonitrile in water containing 0.1% trifluoroacetic acid. Caution: Some proteins may precipitate when concentration of organic solvent is high.
Storage conditions	Short-term (up to a week): Remove buffer, salts and/or acid from column: wash for 10-15 min. using the mobile phase last used, without buffer, salts or acid. For example, if your mobile phase was 50:50 methanol/20 mmol/l phosphate buffer, wash with 50:50 methanol/water. Long-term: Remove buffer, salts and/or acid from column: wash for 10-15 min. using the mobile phase last used, without buffer, salts or acid. Then, replace the solvent with 70% methanol or acetonitrile : 30% water. In either case, tightly plug the column, and store in a cool and dry place.
Usable pH range	1.5-7.5 (pH 2-7.5 recommended)
Max. pressure	20MPa		15MPa
Temperature range	The maximum usable temperature is 60°C. However. for regular use, please use at a constant temperature between 20°C and 50°C.
Usable solvents	Any solvent that will not dissolve the silica gel (such as alkaline solutions) or cleave the stationary phase (such as very acidic solutions) is usable. Note: For solvents with high viscosity, please watch the system pressure and keep it below 20 MPa. Also, please wash the column after using acidic mobile phases or solvents with high freezing points.

Downloads

Brochures

COSMOSIL Protein-R / Wide Pore Columns (2.4 MB)

Ordering Information

COSMOSIL Protein-R

Product Name	Size	Product No.
COSMOSIL Protein-R Guard Column
	4.6 mm l.D. x 10 mm	06518-31
	10 mm l.D. x 20 mm	06528-01
COSMOSIL Protein-R Packed Column
	2.0 mm l.D. x 150 mm	06514-71
	4.6 mm l.D. x 50 mm	06525-31
	4.6 mm l.D. x 150 mm	06526-21
	4.6 mm l.D. x 250 mm	06527-11
	10 mm l.D. x 150 mm	06529-91
	10 mm l.D. x 250 mm	06530-51
	20 mm l.D. x 150 mm	06531-41
	20 mm l.D. x 250 mm	06532-31