COSMOSIL SFC Columns

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.

Conditions
Column	COSMOSIL **	BPR	10 MPa
Column size	4.6 mmI.D. × 250 mm	Temp.	40°C
Column size	4.6 mmI.D. × 250 mm	Detection	UV 273 nm
Mobile phase	A:CO₂B:Methanol B conc. 10%(πMAX only:20%)	Sample	Caffeine (0.10 mg/mL) Theophylline (0.10 mg/mL) Theobromine (0.09 mg/mL)
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.

Conditions
Column	COSMOSIL **	BPR	10 MPa
Column size	4.6 mmI.D. × 250 mm	Temp.	40°C
Column size	4.6 mmI.D. × 250 mm	Detection	UV 254 nm
Mobile phase	A:CO₂B:Methanol B conc. 5%	Sample	o-Hydroxyacetophenone (0.1 mg/mL) m-Hydroxyacetophenone (0.1 mg/mL) p-Hydroxyacetophenone (0.1 mg/mL)
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.

Conditions
Column	COSMOSIL **	BPR	10 MPa
Column size	4.6 mmI.D. × 250 mm	Temp.	40°C
Column size	4.6 mmI.D. × 250 mm	Detection	UV 254 nm
Mobile phase	A:CO₂B: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)
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
Column size	4.6 mmI.D. × 250 mm	Detection	UV 265 nm
Mobile phase	A:CO₂B:Methanol B conc. **%	Sample	Vitamin D₃ (0.6 mg/mL) 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
Column size	4.6 mmI.D. × 250 mm	Detection	UV 260 nm
Mobile phase	A:CO₂B:100 mmol/L Ammonium Acetate -Methanol B conc. **%	Sample	Thymine (0.2 mg/mL) Uracil (0.2 mg/mL) Adenine (0.2 mg/mL) Cytosine (0.2 mg/mL) 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
Column size	4.6 mmI.D. × 250 mm	Detection	UV 220 nm
Mobile phase	A:CO₂B:Tetrahydrofuran B conc. →% 10 min Linear gradient	Sample	Polystyrene, MW906 (5.0 mg/mL) Polystyrene, MW2,200 (5.0 mg/mL)
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
Column size	4.6 mmI.D. × 250 mm	Detection	UV 220 nm
Mobile phase	A:CO₂B:Methanol B conc.2→100% 10min Linear gradient	Sample	Triton X-100 (5.0 mg/mL)
Flow rate	2.0 mL/min	Inj. vol.	5.0 µL

Non-steroidal anti-inflammatory drugs

NSAID chromatogram

Conditions
Column	COSMOSIL HP	Sample	Ibuprofen Fenoprofen Flurbiprofen Ketoprofen 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	Pindolol 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	Inj. Vol.	2.0 µl
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	Inj.Vol.	2.0 µl
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	Inj. Vol.	2.0 µl
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 material	PY	HP	Diol	Cholester	πMAX	PBr
Silica gel	High-purity porous spherical silica
Average particle size (µm)	3, 5		5	5	5	5
Average pore size (Å)	120
Stationary phase
Stationary phase	Pyridinyl	3-Hydroxyphenyl	Diol	Cholesteryl	Pyrenylethyl	Pentabromobenzyl
Endcapping treatment	Endcapped

Downloads

Brochures

COSMOSIL SFC Columns (1.2 MB)

Technical Guide

COSMOSIL SFC Technical Guide (1 MB)

Posters

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

COSMOSIL SFC Columns

Product Information

Selection by Polarity

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

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

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

Applications

Vitamin D

Nucleobases

Polystyrene

Surfactants

Non-steroidal anti-inflammatory drugs

Beta Blockers

Hydrophilic organics

COSMOSIL PY (Pyridinyl)

Fat-Soluble Vitamin Analysis

COSMOSIL Cholester

Column Comparison Data

Amitriptyline

Reserpine

3-Aminobenzoic Acid

Benzoic Acid

Packing Material Properties

Downloads

Brochures

Technical Guide

Posters

Ordering Information

COSMOSIL HP (3-Hydroxyphenyl)

COSMOSIL PY (Pyridinyl)

COSMOSIL Diol

COSMOSIL Cholester

COSMOSIL πMAX

COSMOSIL PBr