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COSMOSIL PBr

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

COSMOSIL PBr
Silica gel High purity porous spherical silica
Average particle size 3 µm            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

Oligonucleotides

 

Polyethylene Glycol (SFC)

 

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

Usage Information

Column5 µm PBr
I.D. (mm)2.03.04.610.020.028.0
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 / References

Brochures
COSMOSIL PBr     
 
Polar molecule separation by COSMOSIL PBr    
     
Posters
Evaluation of Pentabromobenzyl Stationary Phase for Separation of Polar Compounds by Reversed-Phase Chromatography
(Poster from HPLC 2015, Geneva)
 
 
Fat-Soluble and Water-Soluble Vitamins in Functional Water
(Poster from Food Science Asia 2015, Singapore)
 
     
References
  1. Malya, Irine Yunhafita, et al. "Plant growth regulators and Axl and immune checkpoint inhibitors from the edible mushroom Leucopaxillus giganteus." Bioscience, Biotechnology, and Biochemistry (2020): 1-7.
  2. Desmarchelier, Aurélien, Jonas Hamel, and Thierry Delatour. "Sources of overestimation in the analysis of acrylamide-in coffee by liquid chromatography mass spectrometry." Journal of Chromatography A 1610 (2020): 460566.
  3. Chen, Yue, et al. "Anti-inflammatory Limonoids From Cortex Dictamni." Frontiers in Chemistry 8 (2020): 73.
  4. Jiang, Dasen, et al. "Evaluation of a series of phenyl-type stationary phases in supercritical fluid chromatography with the linear solvation energy relationship model and its application to the separation of phenolic compounds." Journal of Chromatography A 1614 (2020): 460700.
  5. Segawa, Hiroki, et al. "Differentiation of ring‐substituted regioisomers of cathinone analogs by supercritical fluid chromatography." Analytical Science Advances.
  6. El-Shaheny, Rania, et al. "Pentabromobenzyl-RP versus triazole-HILIC columns for separation of the polar basic analytes famotidine and famotidone: LC method development combined with in silico tools to follow the potential consequences of famotidine gastric instability." Journal of Pharmaceutical and Biomedical Analysis (2020): 113305.
  7. Yoshii, Katsuhiro, et al. "Exploration of dipeptidyl-peptidase IV (DPP IV) inhibitors in a low-molecular mass extract of the earthworm Eisenia fetida and identification of the inhibitors as amino acids like methionine, leucine, histidine, and isoleucine." Enzyme and Microbial Technology 137 (2020): 109534.
  8. Delatour, Thierry, et al. "Thermal degradation of 2-furoic acid and furfuryl alcohol as pathways in the formation of furan and 2-methylfuran in food." Food chemistry 303 (2020): 125406.

COSMOSIL Application Search

COSMOSIL Application, which includes more than 7,700 applications using COSMOSIL/COSMOCORE columns, is now usable by clicking the link below which is in Nacalai Tesque, Inc. The application is searchable by sample category, sample name, CAS No., column name and particle size

Ordering Information

Product Cat.No. End-Fitting PKG Size Price(US$)  
COSMOSIL 3PBr Packed Column (2.0 mmI.D. x 50 mm ) 19345-91 Waters type 1PKG 620.00 Buy
COSMOSIL 3PBr Packed Column (2.0 mmI.D. x 75 mm ) 19346-81 Waters type 1PKG 620.00 Buy
COSMOSIL 3PBr Packed Column (2.0 mmI.D. x 100 mm ) 19347-71 Waters type 1PKG 620.00 Buy
COSMOSIL 3PBr Packed Column (2.0 mmI.D. x 150 mm ) 19078-01 Waters type 1PKG 620.00 Buy
COSMOSIL 3PBr Packed Column (2.0 mmI.D. x 250 mm ) 19348-61 Waters type 1PKG 740.00 Buy
COSMOSIL 3PBr Packed Column (3.0 mmI.D. x 50 mm ) 19349-51 Waters type 1PKG 620.00 Buy
COSMOSIL 3PBr Packed Column (3.0 mmI.D. x 75 mm ) 19350-11 Waters type 1PKG 620.00 Buy
COSMOSIL 3PBr Packed Column (3.0 mmI.D. x 100 mm ) 19351-01 Waters type 1PKG 620.00 Buy
COSMOSIL 3PBr Packed Column (3.0 mmI.D. x 150 mm ) 19352-91 Waters type 1PKG 620.00 Buy
COSMOSIL 3PBr Packed Column (3.0 mmI.D. x 250 mm ) 19353-81 Waters type 1PKG 740.00 Buy
COSMOSIL 3PBr Packed Column (4.6 mmI.D. x 50 mm ) 19354-71 Waters type 1PKG 620.00 Buy
COSMOSIL 3PBr Packed Column (4.6 mmI.D. x 75 mm ) 19355-61 Waters type 1PKG 620.00 Buy
COSMOSIL 3PBr Packed Column (4.6 mmI.D. x 100 mm) 19356-51 Waters type 1PKG 590.00 Buy
COSMOSIL 3PBr Packed Column (4.6 mmI.D. x 150 mm) 19357-41 Waters type 1PKG 590.00 Buy
COSMOSIL 3PBr Packed Column (4.6 mmI.D. x 250 mm) 19358-31 Waters type 1PKG 690.00 Buy
COSMOSIL 5PBr Packed Column (1.0 mmI.D. x 150 mm) 12881-71 Waters type 1PKG 700.00 Buy
COSMOSIL 5PBr Packed Column (2.0 mmI.D. x 50 mm) 12943-61 Waters type 1PKG 475.00 Buy
COSMOSIL 5PBr Packed Column (2.0 mmI.D. x 100 mm) 13245-81 Waters type 1PKG 500.00 Buy
COSMOSIL 5PBr Packed Column (2.0 mmI.D. x 150 mm) 12392-81 Waters type 1PKG 510.00 Buy
COSMOSIL 5PBr Packed Column (2.0 mmI.D. x 250 mm) 13247-61 Waters type 1PKG 615.00 Buy
COSMOSIL 5PBr Packed Column (3.0 mmI.D. x 50 mm) 12592-61 Waters type 1PKG 535.00 Buy
COSMOSIL 5PBr Packed Column (3.0 mmI.D. x 100 mm) 13249-41 Waters type 1PKG 505.00 Buy
COSMOSIL 5PBr Packed Column (3.0 mmI.D. x 150 mm) 13250-01 Waters type 1PKG 500.00 Buy
COSMOSIL 5PBr Packed Column (3.0 mmI.D. x 250 mm) 13251-91 Waters type 1PKG 540.00 Buy
COSMOSIL 5PBr Packed Column (4.6 mmI.D. x 50 mm) 13252-81 Waters type 1PKG 535.00 Buy
COSMOSIL 5PBr Packed Column (4.6 mmI.D. x 100 mm) 12594-41 Waters type 1PKG 505.00 Buy
COSMOSIL 5PBr Packed Column (4.6 mmI.D. x 150 mm) 12394-61 Waters type 1PKG 475.00 Buy
COSMOSIL 5PBr Packed Column (4.6 mmI.D. x 250 mm) 12395-51 Waters type 1PKG 515.00 Buy
COSMOSIL 5PBr Packed Column (10.0 mmI.D. x 50 mm) 13253-71 Waters type 1PKG 810.00 Buy
COSMOSIL 5PBr Packed Column (10.0 mmI.D. x 100 mm) 13254-61 Waters type 1PKG 940.00 Buy
COSMOSIL 5PBr Packed Column (10.0 mmI.D. x 150 mm) 13255-51 Waters type 1PKG 1,000.00 Buy
COSMOSIL 5PBr Packed Column (10.0 mmI.D. x 250 mm) 12397-31 Waters type 1PKG 1,575.00 Buy
COSMOSIL 5PBr Packed Column (20.0 mmI.D. x 50 mm) 13257-31 Waters type 1PKG 1,450.00 Buy
COSMOSIL 5PBr Packed Column (20.0 mmI.D. x 100 mm) 13258-21 Waters type 1PKG 1,905.00 Buy
COSMOSIL 5PBr Packed Column (20.0 mmI.D. x 150 mm) 13259-11 Waters type 1PKG 2,005.00 Buy
COSMOSIL 5PBr Packed Column (20.0 mmI.D. x 250 mm) 12398-21 Waters type 1PKG 3,675.00 Buy
COSMOSIL 5PBr Packed Column (28.0 mmI.D. x 100 mm) 13260-71 Waters type 1PKG 5,350.00 Buy
COSMOSIL 5PBr Packed Column (28.0 mmI.D. x 150 mm) 13261-61 Waters type 1PKG 5,630.00 Buy
COSMOSIL 5PBr Packed Column (28.0 mmI.D. x 250 mm) 13262-51 Waters type 1PKG 7,550.00 Buy
Guard Column / Guard Cartridge
Product Cat.No. End-Fitting PKG Size Price(US$)  
COSMOSIL 3PBr Guard Cartridge (2.0 mmI.D. x 10 mm ) 19359-34 Waters type 2PKG 220.00 Buy
COSMOSIL 3PBr Guard Cartridge (4.6 mmI.D. x 10 mm ) 19360-94 Waters type 2PKG 220.00 Buy
COSMOSIL 5PBr Guard Cartridge (2.0 mmI.D. x 10 mm) 18196-94 Waters type 2PKG 220.00 Buy
COSMOSIL 5PBr Guard Cartridge (4.6 mmI.D. x 10 mm) 12444-14 Waters type 2PKG 230.00 Buy
COSMOSIL 5PBr Guard Column (2.0 mmI.D. x 10 mm) 13287-41 Waters type 1PKG 450.00 Buy
COSMOSIL 5PBr Guard Column (10.0 mmI.D. x 20 mm) 12396-41 Waters type 1PKG 555.00 Buy
COSMOSIL 5PBr Guard Column (20.0 mmI.D. x 20 mm) 13256-41 Waters type 1PKG 1,200.00 Buy
COSMOSIL 5PBr Guard Column (28.0 mmI.D. x 50 mm) 12653-61 Waters type 1PKG 3,455.00 Buy
HPLC Accessories
Product Cat.No. End-Fitting PKG Size Price(US$)  
COSMOSIL Direct Cartridge Holder 19989-71 Waters type 1 pkg 270.00 Buy
COSMOSIL Guard Cartridge Holder (4.6 mm I.D.) 38009-79 Waters type 1 pkg 265.00 Buy
COSMOSIL Guard Cartridge Holder (2.0 mm I.D.) 11884-71 Waters type 1 pkg 265.00 Buy
COSMOSIL Column Prefilter 39361-19 Waters type 1 pkg 145.00 Buy
COSMOSIL Column Spare Filter for Prefilter (1 micron) 39539-09 Waters type 2 pkg 55.00 Buy
COSMOSIL Column Connecting Tube (0.25 mm) 37843-69 Waters type 1 pkg 65.00 Buy
COSMOSIL Column Connecting Tube (0.1 mm) 12570-41 Waters type 1 pkg 65.00 Buy