Products

COSMOSIL SFC Columns

Column Image

Supercritical Fluid Chromatography (SFC) has become more attractive because it offers some advantages over HPLC, such as high speed, unique selectivity and environmentally friendly separations. Many conventional normal-phase stationary phases, such as diol, amino and cyano, have been used for SFC applications. However, these phases present limitations for separations. COSMOSIL SFC Columns have been developed to enhance the capability of SFC separations.

Features:

  • Three categories (I–III) of stationary phase for different types of compounds
  • Different selectivity from HPLC

>> Read our paper in Molecules (open access)

Product Information

Category I: Columns for mid- to high-polarity compounds

For these compounds, a high-polarity stationary phase is suitable. More polar compounds are retained longer.

Product Stationary phase Features
COSMOSIL PY Pyridinyl Similar selectivity to 2-ethylpyridine; strong retention in general.
COSMOSIL HP 3-Hydroxyphenyl Different selectivity from PY; strong retention for basic compounds.
COSMOSIL Diol Diol Less effect from ionic interaction.

Category II: Columns for low-polarity compounds

For these compounds, a low-polarity stationary phase is suitable.

Product Stationary phase Features
COSMOSIL Cholester Cholesteryl Longer retention and better separation than C18.

Category III: Columns for SFC-specific separations

In supercritical fluid chromatography (SFC), secondary interactions such as π-π and dispersion force* are stronger compared to high-performance liquid chromatography (HPLC). As a result, these columns are capable of unique separations in SFC.

Product Stationary phase Features
COSMOSIL πMAX Pyrenylethyl Stronger π-π interactions than phenyl columns.
COSMOSIL PBr Pentabromobenzyl Unique separations using dispersion force.*

* Dispersion 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 of whether they are polar or non-polar. Compounds with high polarizability have stronger dispersion force.

Selection by Polarity

Derivatives of xanthine (high-polarity compounds) XLogP3: -0.8 – 0.0

Category I columns (COSMOSIL 5PY, 5HP, 5Diol) and COSMOSIL 5πMAX separated the sample well. The elution order with COSMOSIL 5πMAX was different than the category I columns.

xanthine derivatives chromatogram

Conditions
Column COSMOSIL ** BPR 10 MPa
Column size 4.6 mmI.D. × 250 mm Temp. 40°C
Detection UV 273 nm
Mobile phase A:CO2
B:Methanol
B conc. 10%(πMAX only:20%)
Sample
  1. Caffeine (0.10 mg/mL)
  2. Theophylline (0.10 mg/mL)
  3. Theobromine (0.09 mg/mL)
  4. sfc-1-sample.jpg

Flow rate 3.0 mL/min Inj. vol. 5.0 µL

Positional isomers (mid-polarity compounds) XLogP3: 1.4 – 1.9

Category I columns (COSMOSIL 5PY, 5HP, 5Diol) yielded the best separation.

positional isomers chromatogram 1

Conditions
Column COSMOSIL ** BPR 10 MPa
Column size 4.6 mmI.D. × 250 mm Temp. 40°C
Detection UV 254 nm
Mobile phase A:CO2
B:Methanol
B conc. 5%
Sample

o-Hydroxyacetophenone (0.1 mg/mL)

m-Hydroxyacetophenone (0.1 mg/mL)

p-Hydroxyacetophenone (0.1 mg/mL)

sfc-2-sample.jpg

Flow rate 3.0 mL/min Inj. vol. 5.0 µL

Positional isomers (low-polarity compounds) XLogP3: 5.8 – 6.0

Category III (COSMOSIL 5πMAX, 5PBr) and category II (5Cholester) columns separated most effectively.

positional isomers chromatogram 2

Conditions
Column COSMOSIL ** BPR 10 MPa
Column size 4.6 mmI.D. × 250 mm Temp. 40°C
Detection UV 254 nm
Mobile phase A:CO2
B:Methanol
B conc. 5%(PBr only: 15%)
Sample

o-Terphenyl(0.1 mg/mL)

m-Terphenyl(0.1 mg/mL)

p-Terphenyl(0.1 mg/mL)


sfc-3-sample.jpg
Flow rate 3.0 mL/min Inj. vol. 5.0 µL

Applications

Vitamin D

COSMOSIL 5πMAX and 5PBr were able to separate the vitamins. The PFP (pentafluorophenyl) column, which also uses a halogenated stationary phase, could not separate them. The dispersion force used by PBr interacts more strongly with larger molecules. PFP, which is a smaller molecule, does not exhibit this selectivity.

vit. d chromatogram

Conditions
Column COSMOSIL ** BPR 10 MPa
Column size 4.6 mmI.D. × 250 mm Temp. 40°C
Detection UV 265 nm
Mobile phase A:CO2
B:Methanol
B conc. **%
Sample
  1. Vitamin D3 (0.6 mg/mL)
  2. Vitamin D2 (0.6 mg/mL)
Flow rate 3.0 mL/min Inj. vol. 3.0 µL

Nucleobases

COSMOSIL 5Diol separated 5 nucleic acid bases.

nucleobase chromatogram

Conditions
Column COSMOSIL ** BPR 10 MPa
Column size 4.6 mmI.D. × 250 mm Temp. 40°C
Detection UV 260 nm
Mobile phase A:CO2
B:100 mmol/L Ammonium Acetate -Methanol
B conc. **%
Sample
  1. Thymine (0.2 mg/mL)
  2. Uracil (0.2 mg/mL)
  3. Adenine (0.2 mg/mL)
  4. Cytosine (0.2 mg/mL)
  5. Guanine (0.2 mg/mL)
Flow rate 3.0 mL/min Inj. vol. 5.0 µL

Polystyrene

These polystyrene samples with different degrees of polymerization are likely separated by number of monomer units. COSMOSIL 5πMAX and 5PBr were able to separate the high-MW polystyrene.

polystyrene chromatogram

Conditions
Column COSMOSIL ** BPR 10 MPa
Column size 4.6 mmI.D. × 250 mm Temp. 40°C
Detection UV 220 nm
Mobile phase A:CO2
B:Tetrahydrofuran
B conc. *→*% 10 min Linear gradient
Sample

Polystyrene, MW906 (5.0 mg/mL)

Polystyrene, MW2,200 (5.0 mg/mL)

sfc-6-sample.jpg

Flow rate 3.0 mL/min Inj. vol. 5.0 µL

Surfactants

surfactant chromatogram

Conditions
Column COSMOSIL 5PBr BPR 10 MPa
Column size 4.6 mmI.D. × 250 mm Temp. 40°C
Detection UV 220 nm
Mobile phase A:CO2
B:Methanol
B conc.2→100% 10min Linear gradient
Sample

Triton X-100 (5.0 mg/mL)

sfc-8-sample.jpg

Flow rate 2.0 mL/min Inj. vol. 5.0 µL

Non-steroidal anti-inflammatory drugs

NSAID chromatogram

Conditions
Column COSMOSIL HP Sample
  1. Ibuprofen
  2. Fenoprofen
  3. Flurbiprofen
  4. Ketoprofen
  5. Indoprofen
Column size 4.6 mm I.D. x 150 mm
Mobile phase Carbon Dioxide : Methanol
Gradient 95:5 to 65:35 over 6 minutes
Flow rate 5.0 ml/min
Temperature 30°C
Detection UV 230 nm

Beta Blockers

Peak elution order reversed under identical conditions.

beta blocker chromatogram

Conditions
Column COSMOSIL HP and a 2-ethylpyridine column Sample
  1. Pindolol
  2. Atenolol
Column size 4.6 mm I.D. x 150 mm
Mobile phase Carbon Dioxide : Methanol
w/20 mM ammonium formate
Gradient 95:5 to 50:50 over 4 minutes
Flow rate 5.0 ml/min
Temperature 30°C
Detection APCI(+)

Hydrophilic organics

COSMOSIL PY (Pyridinyl)

hydrophilic organics chromatogram

Conditions
Column size 4.6 mm I.D. x 150 mm
Flow rate 5.6 mL/min
Pressure 140 bar
Temperature Ambient
Mobile phase Methanol gradient 5-50%
@18%/minute;
hold @ 50% 0.1 min;
return to 5% @99%/min

Fat-Soluble Vitamin Analysis

COSMOSIL Cholester

When used with SFC, COSMOSIL Cholester can separate fat-soluble vitamins and their impurities.

fat-soluble vitamin chromatogram

Conditions
Column size 4.6 mm I.D. x 250 mm
Mobile phase IPA in CO2
IPA conc. 0→10%(0–10min),10%(10–12min)
Flow rate 3.0 ml/min
BPR 15 MPa
Temperature 40°C
Detection UV-VIS
Sample conc 2.0 mg/ml
Inj. Vol 1.0 µl
Sample

COSMOSIL Cholester exhibits strong retention for fat-soluble vitamins and is suitable for on-line SFE-SFC using Shimadzu’s Nexera UC. The on-line extraction from food also produced triglyceride impurities, which were successfully separated from the
vitamins.
Data courtesy of Shimadzu Corporation

Column Comparison Data

Amitriptyline

Conditions
Column size 2.1 mm I.D. × 150 mm Temperature 40°C
Mobile phase A: CO2
B: 0.1% CH3COONH4 -Methanol
B conc. 0→60% (0–14 min), 60% (14–17 min)
Detection UV 250 nm
Inj.Vol.: 2.0 µl
Sample Amitriptyline (1 mmol/l)
Flow rate 0.8 ml/min
BPR 10 MPa

Data courtesy of Kyushu University Medical Institute of Bioregulation Research Center for Transomics Medicine Division of Metabolomics

Reserpine

Conditions
Column size 2.1 mm I.D. × 150 mm BPR 10 MPa
Mobile phase A: CO2
B: 0.1% CH3COONH4 -Methanol
B conc. 0→60% (0–14 min), 60% (14–17 min)
Temperature 40°C
Detection UV 260 nm
Inj. Vol. 2.0 µl
Flow rate 0.8 ml/min
Sample Reserpine (1mmol/l)

Data courtesy of Kyushu University Medical Institute of Bioregulation Research Center for Transomics Medicine Division of Metabolomics

3-Aminobenzoic Acid

Conditions
Column size 2.1 mm I.D. × 150 mm BPR 10 MPa
Mobile phase A: CO2
B: 0.1% CH3COONH4 -Methanol
B conc. 0→60% (0–14 min), 60% (14–17 min)
Temperature 40°C
Detection UV 250 nm
Inj.Vol. 2.0 µl
Flow rate 0.8 ml/min
Sample 3-Aminobenzoic Acid (10mmol/l)

Data courtesy of Kyushu University Medical Institute of Bioregulation Research Center for Transomics Medicine Division of Metabolomics

Benzoic Acid

Conditions
Column size 2.1 mm I.D. × 150 mm BPR 10 MPa
Mobile phase A: CO2
B: 0.1% CH3COONH4 -Methanol
B conc. 0→60% (0–14 min), 60% (14–17 min)
Temperature 40°C
Detection UV 230nm
Inj. Vol. 2.0 µl
Flow rate 0.8 ml/min
Sample Benzoic Acid (10 mmol/l)

Data courtesy of Kyushu University Medical Institute of Bioregulation Research Center for Transomics Medicine Division of Metabolomics

See more COSMOSIL Applications

Packing Material Properties

Packing materialPYHPDiolCholesterπMAXPBr
Silica gel High-purity porous spherical silica
Average particle size (µm) 3, 5 5 5 5 5
Average pore size (Å) 120
Stationary phase PY-kouzousiki.png paiMAX-kouzousiki.jpg
Pyridinyl 3-Hydroxyphenyl Diol Cholesteryl Pyrenylethyl Pentabromobenzyl
Endcapping treatment Endcapped

Downloads

Brochures

Technical Guide

  • COSMOSIL SFC Technical Guidepdf (1 MB)

Posters

  • Research into the Retention Mechanism of SFC
    (Poster from SFC Asia 2017, Osaka, Japan)
    pdf (358 KB)
  • Development of novel stationary phases for Supercritical Fluid Chromatography (SFC) (Poster from HPLC 2011, Budapest, Hungary)
    pdf (242 KB)

Ordering Information

COSMOSIL HP (3-Hydroxyphenyl)

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COSMOSIL PY (Pyridinyl)

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COSMOSIL Diol

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COSMOSIL Cholester

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COSMOSIL πMAX

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COSMOSIL PBr

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