Products

COSMOSIL PBr

Features

Column Image

  • Separate hydrophilic compounds in reversed-phase conditions
  • Simple mobile phase condition compared to HILIC
  • More sample loading capacity than HILIC
  • Alternative selectivity to C18 column

Product Information

Specifications

Packing Material 5PBr
Silica Gel High purity porous spherical silica
Average Particle Size 5 µm
Average Pore Size approx. 120 Å
Specific Surface Area approx. 300 m2/g
Stationary Phase Stationary Phase
Pentabromobenzyl group
Bonding Type Monomeric
Endcapping treatment Yes
Carbon load approx. 8%
Usable pH range 2-7.5

Separation of hydrophilic compounds in reversed-phase conditions

Hydrophilic interaction chromatography (HILIC) is an increasingly popular analytical technique. However, it is often difficult to develop a robust method due to users’ less familiarity with HILIC conditions. Furthermore, high concentration of acetonitrile used in HILIC mobile phase makes it extremely sensitive to samples’ water concentration; injecting samples in high water concentration often results in poor peak shapes. COSMOSIL PBr enables separation of hydrophilic compounds in reversedphase conditions, maintaining sharp peak shapes even with aqueous samples.

Chromatogram

Condition
Column size 4.6 mm I.D. x 150 mm Sample
  • Uracil (0.1 mg/ml)
  • Uridine (0.2 mg/ml)
Mobile phase HILIC: Acetonitrile / H2O = 90/10
PBr:H2O
Flow Rate 1.0 ml/min Sam. solution H2O
Temperature 30°C Structure
Detection UV260nm

Comparison with C18 Columns

COSMOSIL PBr retains hydrophilic compounds stronger than C18 columns under the same reversed-phase conditions.

Chromatogram

Condition
Column size 4.6 mm I.D.×250 mm Sample
  1. Ammelide (0.1 mg/ml)
  2. Cyanuric Acid (0.5 mg/ml)
  3. Ammeline (0.1 mg/ml)
  4. Melamine (0.1 mg/ml)
Structure
Mobile phase 20 mmol/l Phosphate Buffer(pH7.0)
Flow Rate 1.0 ml/min
Temperature 30°C
Detection UV225nm Inj. Vol. 1.0 ul

Separation Mechanism

In reversed-phase chromatography, compounds are separated by difference in hydrophobicity (the number of CH2 bases, see chromatogram on the left (1)). Compounds with little hydrophobicity (the number of OCH2CH2, see chromatogram on the right (2)), are not retained by a C18 column. However, these compounds can easily be separated by dispersion force interaction using the COSMOSIL PBr column.

Compounds with more hydrophobicity

Chromatogram

Condition
Column size 4.6 mm I.D.×150 mm Sample
  • Benzene
  • Toluene
  • Ethylbenzene
  • n-Propylbenzene
  • n-Butylbenzene
Mobile phase Methanol/ H2O = 80/20
Flow rate 1.0 ml/min
Temperature 30°C Structure
Detection UV254nm

Compounds with less hydrophobicity

Chromatogram

Condition
Column size 4.6 mm I.D.×150 mm Sample
  • Benzyl Alcohol
  • Ethylene Glycol Monobenzyl Ether
  • Diethylene Glycol Monobenzyl Ether
  • Triethylene Glycol Monobenzyl Ether
  • Tetraethylene Glycol Monobenzyl Ether
Mobile phase Methanol/ H2O = 60/40
Flow rate 1.0 ml/min
Temperature 30°C
Detection UV254nm

Dispersion force (instantaneous dipole?induced dipole force)

London Dispersion force is a weak intermolecular force that results from dipoles temporarily induced from random unsymmetrical electron positions in two adjacent atoms, also known as “instantaneous dipole? induced dipole force”. It is present in all molecules regardless if they are polar or non-polar. Compounds with high polarizability have stronger dispersion force.

Compounds with stronger dispersion force

  • Larger and heavier molecules
  • Molecules with larger and heavier atoms (e.g. from weakest to strongest in halogens, F2, Cl2, Br2, and I2)
  • Molecules with delocalized electrons and resonance (e.g. aromatic compounds)

COSMOSIL PBr column is packed with pentabromobenzyl bonded silica that enables separation by dispersion force interaction.

Suitable Compounds to Analyze by COSMOSIL PBr

COSMOSIL PBr retains cyclic compounds or amide compounds more strongly than C18 columns.

(Mobile phase)
(1) Methanol/ 20mmol/l Phosphate buffer(pH7.0) = 10/90
(2) 20mmol/l Phosphate buffer(pH2.5)
(3) 20mmol/l Phosphate buffer(pH7.0)
(4) Methanol/ H2O = 10/90

Column Stability Test

COSMOSIL PBr column remains stable after 1000 injections under 100% aqueous condition.

Condition
Column size 4.6 mm I.D. × 150 mm Sample
  • L-Noradrenaline (0.5 mg/ml)
  • L-Adrenaline (0.5 mg/ml)
  • Dopamine (0.5 mg/ml)
  • L-DOPA (0.5 mg/ml)
Mobile phase 20 mmol/l Phosphate buffer(pH2.5)
Flow rate 1.0 ml/min
Temperature 40°C
Detection UV280nm Inj.Vol. 1.0 µl

Applications

Raffinose and Stachyose

Application

Condition
Column size 4.6 mm I.D. x 250 mm Sample 1; Raffinose (5 mg/ml)
2; Stachyose (5 mg/ml)
Mobile phase Methanol/ H2O = 5/95
Flow rate 1.0 ml/min
Temperature 30°C
Detection RI Inj.Vol. 2.0 µl

Gentamycin

Condition
Column size 4.6 mm I.D. x 150 mm Sample Gentamicin
(C1a, C2, C2a, C1)
Mobile phase 0.1% Pentafluoropropionic Acid -Acetonitrile/ H2O = 10/90
Flow rate 0.2 ml/min
Temperature 40°C
Detection ESI-MS, Positive, SIM Inj.Vol. 1.0 µl

Nucleic Acid Base

Condition
Column Size 4.6 mm I.D. x 150 mm
Mobile Phase Methanol/ 20mmol/l Phosphate buffer(pH7.0) = 10/90
Flow Rate 1.0 ml/min
Temperature 30°C
Detection UV260nm
Sample
  1. Cytosine (0.05 mg/ml)
  2. Uracil (0.05 mg/ml)
  3. Thymine (0.05 mg/ml)
  4. Guanine (0.05 mg/ml)
  5. Adenine (0.05 mg/ml)
Inj.Vol. 1.0 µl

Coenzyme A

Condition
Column Size 4.6 mm I.D. x 150 mm
Mobile Phase Methanol/ 20mmol/l Phosphate buffer(pH7.0) = 20/80
Flow Rate 1.0 ml/min
Temperature 30°C
Detection UV260nm
Sample
  1. Malonyl Coenzyme A (0.25 mg/ml)
  2. Coenzyme A (0.25 mg/ml)
  3. Acetyl Coenzyme A (0.25 mg/ml)
Inj.Vol. 2.0 µl

Methylimidazole Isomers

Condition
Column Size 4.6 mm I.D. x 250 mm
Mobile Phase 20mmol/l Phosphate buffer(pH2.5)
Flow Rate 1.0 ml/min
Temperature 30°C
Detection UV220nm
Sample
  1. 1-Methylimidazole (0.1 mg/ml)
  2. 2-Methylimidazole (0.1 mg/ml)
  3. 4-Methylimidazole (0.1 mg/ml)
Inj.Vol. 1.0 µl

Applications

Surfactants

COSMOSIL PBr shows excellent separation for surfactants compared to C18 or PFP columns.

Condition
Column size 4.6 mm I.D. x 150 mm Sample Triton(R) X-100 (50 mg/ml)
Mobile phase 5C18-MS-II, 5PFP: Methanol/ H2O = 80/20
PBr: Methanol
Flow rate 1.0 ml/min
Temperature 30°C
Detection UV254nm Inj.Vol. 2.0 µl

Halogen Compounds

COSMOSIL PBr shows high selectivity for halogen compounds.

Condition
Column size 4.6 mm I.D. x 150 mm Sample
  1. Fluorobenzene (1.0 mg/ml)
  2. α,α,α-Trifluorotoluene
    [Benzotrifluoride] (0.25 mg/ml)
  3. Toluene (2.5 mg/ml)
  4. Chlorobenzene (5.0 mg/ml)
  5. Bromobenzene (5.0 mg/ml)
  6. Iodobenzene (2.0 mg/ml)
Mobile phase Methanol/ H2O = 65/35
Flow rate 1.0 ml/min
Temperature 30°C
Detection UV254nm Inj.Vol. 1.0 µl

See more COSMOSIL Applications

Usage information

Column 5 µm PBr
I.D. (mm) 2.0 3.0 4.6 10.0 20.0 28.0
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. For example, if your mobile phase was 50:50 methanol/20 mmol/l phosphate buffer, wash with 50:50 methanol/water.

2. (If you used a buffer, always do step 1 first!) Remove adsorbed compounds and fix unstable baselines by washing with methanol and/or THF.
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 pH2~pH7.5
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.
Other instructions - Buffer concentration is usually sufficient at 0.005 - 0.02 mol/L.
- Always filter mobile phases using a 0.5 µm or finer filter before use.
- Acetonitrile is not recommended as a mobile phase.

Downloads

Brochures

  • COSMOSIL PBr pdf (1.4 MB)
  • Polar molecule separation by COSMOSIL PBrpdf(1.7 MB)
  • LC-MS/MS Analysis of Monoamines in Mouse Brain Tissue pdf(1 MB)

Posters

  • Evaluation of Pentabromobenzyl Stationary Phase for Separation of Polar Compounds by Reversed-Phase Chromatography
    (Poster from HPLC 2015, Geneva)
    pdf (370 KB)
  • Fat-Soluble and Water-Soluble Vitamins in Functional Water
    (Poster from Food Science Asia 2015, Singapore)
    pdf (870 KB)

Ordering Information

Analytical and Preparative Column (Particle size : 5 µm)

COSMOSIL 5PBr

Product Name Size Product No. Inquiry
COSMOSIL 5PBr
Guard Column
4.6 mm I.D. x 10 mm
cartridge*
12444-14 Inquire
Now
10.0 mm I.D. x 20.0 mm 12396-41
20.0 mm I.D. x 20.0 mm 13256-41
COSMOSIL Guard
Cartridge Holder
4.6 mm I.D. 38009-79
COSMOSIL PBr
Column
2.0 mm l.D. x 50 mm 12943-61 Inquire
Now
2.0 mm l.D. x 100 mm 13245-81
2.0 mm l.D. x 150 mm 12392-81
2.0 mm l.D. x 250 mm 13247-61
3.0 mm l.D. x 50 mm 12592-61
3.0 mm l.D. x 100 mm 13249-41
3.0 mm l.D. x 150 mm 13250-01
3.0 mm l.D. x 250 mm 13251-91
4.6 mm l.D. x 50 mm 13252-81
4.6 mm l.D. x 150 mm 12394-61
4.6 mm l.D. x 250 mm 12395-51
10.0 mm I.D. x 50 mm 13253-71
10.0 mm I.D. x 100 mm 13254-61
10.0 mm I.D. x 150 mm 13255-51
10.0 mm I.D. x 250 mm 12397-31
20.0 mm I.D. x 50 mm 13257-31
20.0 mm I.D. x 100 mm 13258-21
20.0 mm I.D. x 150 mm 13259-11
20.0 mm I.D. x 250 mm 12398-21
28.0 mm I.D. x 100 mm 13260-71
28.0 mm I.D. x 150 mm 13261-61
28.0 mm I.D. x 250 mm 13262-51

*Cartridge Holder is required.

(Storage) RT: Room temperature, A: Cool and dark, R: Refrigerator, F: Freeze