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COSMOSIL 5PBr

Separation of hydrophilic compounds in reversed-phase conditions

  • 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

Packing MaterialCOSMOSIL 5PBr
Silica gel High purity porous spherical silica
Average particle size 5 µm
Average pore size approx. 120Å
Specific surface area approx. 300 m²/g
Stationary phase Pentabromobenzyl group
Bonding type Monomeric
Endcapping treatment Yes
Carbon content 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 reversed phase conditions, maintaining sharp peak shapes even with aqueous samples.

Column size: 4.6 mmI.D. x 150 mm
Mobile phase: HILIC; Acetonitrile : Water = 90 : 10
PBr; Water
Flow rate: 1.0 ml/min
Temperature: 30°C
Detection: UV 260 nm
Sample: 1. Uracil (0.1 mg/ml)
2. Uridine (0.2 mg/ml)
Sample solution: Water

Comparison with C18 Columns

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

Column size: 4.6 mmI.D. x 250 mm
Mobile phase: 20 mmol/l Phosphate Buffer (pH7.0)
Flow rate: 1.0 ml/min
Temperature: 30°C
Detection: UV 225 nm
Sample: 1. Ammelide (0.1 mg/ml)
2. Cyanuric Acid (0.5 mg/ml)
3. Ammeline (0.1 mg/ml)
4. Malamine (0.1 mg/ml)
Injection vol.: 1.0 µl

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

Column size: 4.6 mmI.D. x 150 mm
Mobile phase: Methanol : Water = 80 : 20
Flow rate: 1.0 ml/min
Temperature: 30°C
Detection: UV 254 nm
Sample: 1. Benezene
2. Toluene
3. Ethylbenzene
4. n-Propylbenzene
5. n-Butylbenzene

Compounds with less hydrophobicity

Column size: 4.6 mmI.D. x 150 mm
Mobile phase: Methanol : Water = 60 : 40
Flow rate: 1.0 ml/min
Temperature: 30°C
Detection: UV 254 nm
Sample: 1. Benzyl Alcohol
2. Ethylene Glycol Monobenzyl Ether
3. Diethylene Glycol Monobenzyl Ether
4. Triethylene Glycol Monobenzyl Ether
5. Tetraethylene Glycol Monobenzyl Ether

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 : 20 mmol/l Phosphate buffer(pH7.0) = 10 : 90)
(2) 20 mmol/l Phosphate buffer (pH2.5))
(3) 20 mmol/l Phosphate buffer (pH7.0))
(4) Methanol : Water = 10 : 90

Column Stability Test

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

Column size: 4.6 mmI.D. x 150 mm
Mobile phase: 20 mmol/l Phosphate buffer (pH2.5)
Flow rate: 1.0 ml/min
Temperature: 40°C
Detection: UV 280 nm
Sample: 1. L-Noradrenaline (0.5 mg/ml)
2. L-Adrenaline (0.5 mg/ml)
3. Dopamine (0.5 mg/ml)
4. L-DOPA (0.5 mg/ml)
Injection vol.: 1.0 µl

Application

Gentamicin

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

Nucleic Acid Base

Column size: 4.6 mmI.D. x 150 mm
Mobile phase: Methanol : 20 mmol/l Phosphate buffer(pH7.0) = 10 : 90
Flow rate: 1.0 ml/min
Temperature: 30°C
Detection: UV 260 nm
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)
Injection vol.: 1.0 µl

Coenzyme A

Column size: 4.6 mmI.D. x 150 mm
Mobile phase: Methanol : 20 mmol/l Phosphate buffer(pH7.0) = 20 : 80
Flow rate: 1.0 ml/min
Temperature: 30°C
Detection: UV 260 nm
Sample: 1. Malonyl Coenzyme A (0.25 mg/ml)
2. Coenzyme A (0.25 mg/ml)
3. Acetyl Coenzyme A (0.25 mg/ml)
Injection vol.: 2.0 µl

Methylimidazole Isomers

Column size: 4.6 mmI.D. x 150 mm
Mobile phase: 20 mmol/l Phosphate buffer(pH2.5)
Flow rate: 1.0 ml/min
Temperature: 30°C
Detection: UV 220 nm
Sample: 1. 1-Methylimidazole (0.1 mg/ml)
2. 2-Methylimidazole (0.1 mg/ml)
3. 4-Methylimidazole (0.1 mg/ml)
Injection vol.: 1.0 µl

Surfactants

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

Column size: 4.6 mmI.D. x 150 mm
Mobile phase: 5C18-MS-II, 5PFP; Methanol : Water = 80 : 20
PBr; Methanol
Flow rate: 1.0 ml/min
Temperature: 30°C
Detection: UV 254 nm
Sample: Triton® X-100 (50 mg/ml)
Injection vol.: 2.0 µl

Halogen Compounds

Column size: 4.6 mmI.D. x 150 mm
Mobile phase: Methanol : Water = 65 : 35
Flow rate: 1.0 ml/min
Temperature: 30°C
Detection: UV 254 nm
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)
Injection vol.: 1.0 µl

Ordering Information

Analytical
Product Cat.No. End-Fitting Quantity Price(US$)  
COSMOSIL PBr Packed Column, 2.0 mm I.D. x 50 mm 12943-61 Waters type 1 pkg 450.00 Buy
COSMOSIL PBr Packed Column, 2.0 mm I.D. x 150 mm 12392-81 Waters type 1 pkg 600.00 Buy
COSMOSIL PBr Packed Column, 3.0 mm I.D. x 50 mm 12592-61 Waters type 1 pkg 350.00 Buy
COSMOSIL PBr Packed Column, 4.6 mm I.D. x 100 mm 12594-41 Waters type 1 pkg 480.00 Buy
COSMOSIL PBr Packed Column, 4.6 mm I.D. x 150 mm 12394-61 Waters type 1 pkg 500.00 Buy
COSMOSIL PBr Packed Column, 4.6 mm I.D. x 250 mm 12395-51 Waters type 1 pkg 630.00 Buy
Guard Cartridge/Column
Product Cat.No. End-Fitting Quantity Price(US$)  
COSMOSIL PBr Guard Cartridge, 4.6 mm I.D. x 10 mm 12444-14 Waters type 2 pkg 220.00 Buy
COSMOSIL Guard Cartridge Holder (4.6 mm I.D.) 38009-79 Waters type 1 pkg 250.00 Buy
COSMOSIL PBr Guard Column, 10.0 mm I.D. x 20 mm 12396-41 Waters type 1 pkg 530.00 Buy
COSMOSIL PBr Guard Column, 20.0 mm I.D. x 20 mm 13256-41 Waters type 1 pkg 1,150.00 Buy
Preparative
Product Cat.No. End-Fitting Quantity Price(US$)  
COSMOSIL PBr Packed Column, 10.0 mm I.D. x 250 mm 12397-31 Waters type 1 pkg 1,500.00 Buy
COSMOSIL PBr Packed Column, 20.0 mm I.D. x 250 mm 12398-21 Waters type 1 pkg 3,500.00 Buy