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Supercritical Fluid Chromatography
In supercritical fluid chromatography (SFC), a substance in a supercritical state is used as the mobile phase. SFC is particularly effective in the separation of lipophilic and medium-polarity compounds and offers an environmentally friendly alternative to traditional chromatographic techniques, which often rely on the use of large quantities of organic solvents. Supercritical CO₂, which is most commonly used as the mobile phase in SFC, is not only inexpensive and non-toxic, but also easy to remove from the analysed substances, making the technique particularly attractive for the food, pharmaceutical and chemical industries.
Despite its many advantages, the application of SFC is not without its challenges. The need for specific equipment that can handle the high pressures and temperatures and the development of suitable methods for specific analytes can limit the implementation of the technique in some laboratories. Nevertheless, with advancing technology and a growing recognition of its environmentally friendly benefits, SFC is gaining popularity and is seen as a valuable addition to existing chromatographic techniques.
Technical Data
Basics
What does supercritical state mean?
The supercritical state describes a region in the phase diagram in which the respective compound is present neither as a gas nor as a liquid, but only as a phase, which is referred to as fluid or supercritical. This area is defined by the so-called critical temperature (Tc) and the critical pressure (pc). If these are reached or exceeded for a particular substance, it is in a supercritical state.
Which liquids or gases can be used as a mobile phase?
As carbon dioxide (CO2) can be brought to the supercritical state relatively easily (Tc ≈ 31 °C, pc ≈ 73 bar) and is available in almost unlimited quantities, it is usually used as a mobile phase. However, other compounds can also be used in principle. The only requirement for this is that the mobile phases used cannot be converted to the supercritical state with too much effort. In order to change the elution strength of the mobile phase, certain modifiers are added, e.g. methanol or ethanol.
And which stationary phases are used?
Theoretically, all silica- and polymer-based stationary phases can be used in SFC, whereby the stationary phases should always be selected depending on the respective application. It should be noted that SFC behaves similarly to normal phase chromatography, so that stationary phases are often used that are also used in normal phase chromatography, e.g. pure silica, diol, cyano, HILIC or amino phases. Pyridine, 2- and 4-ethylpyridine, 2-picolylamine or aminoanthracene phases are also used, which were specially developed for SFC and are said to be particularly suitable for basic analytes. In addition to pressure-stable polymer-based phases, "normal" C18, C8, phenyl or pentafluorophenyl phases can also be used. Most chiral stationary phases can be used for the separation of enantiomers.
What special equipment is required for SFC?
The equipment used for SFC is very similar to conventional HPLC systems. They simply need to be able to control the temperature and pressure precisely, from the sample feed to the detector. This means that the detector cell must be pressure-stable, at least one pump must be able to deliver liquid CO2 reproducibly and constantly and a back-pressure regulator must be available which, together with the column oven, maintains the required physical conditions. There are now manufacturers who sell complete SFC devices as well as those who offer special SFC kits that can be used to make existing HPLC systems SFC-compatible.
Applications
Where is the SFC used?
There are many applications for SFC, e.g. in the analysis of drugs, food, explosives, petroleum or polymers. SFC can also be used to separate enantiomers from chiral phases, which is one of the main applications of SFC today. Separations on a preparative scale for product isolation are also possible. In general, SFC is becoming increasingly popular as a separation method because a large number of high-quality columns and reliable systems are now available. Furthermore, SFC allows large quantities of organic solvents to be saved, which is why SFC is also referred to as "green chemistry".
Downloads
Manufacturer of special SFC phases
| Manufacturer | Name | Modification | Pore size | Particle size |
| Chromanik | Sunshell 2-EP | 2-Ethylpyridine | 90 Å | 2.6 µm |
| Princeton Chromatography | 2-ethylpyridines | 2-Ethylpyridines | 60 Å | 3, 5, & 10 µm |
Silica | - | 60 Å | 3, 5, & 10 µm | |
Cyano | Cyanopropyl | 60 Å | 3, 5, & 10 µm | |
DIOL | Diol | 60 Å | 3, 5, & 10 µm | |
DIOL-HL | Diol | 60 Å | 5 & 10 µm | |
2CN:DIOL | Cyanopropyl & diol | 60 Å | 3, 5, & 10 µm | |
Amino | Aminopropyl | 60 Å | 3, 5, & 10 µm | |
DEAP | Diethylaminopropyl | 60 Å | 3, 5, & 10 µm | |
benzamides | Propyl benzamides | 100 Å | 3, 5, & 10 µm | |
PA | Propylbenzamides | 60 Å | 3, 5, & 10 µm | |
PPU | Propylpyridylurea | 100 Å | 3, 5, & 10 µm | |
Propylurea | Propylurea | 100 Å | 3, 5, & 10 µm | |
DNP | Dinitrophenyl | 100 Å | 3, 5, & 10 µm | |
Pyridine amides | Pyridine amides | 60 Å | 3, 5, & 10 µm | |
4-Ethylpyridines | 4-Ethylpyridines | 60 Å | 3, 5, & 10 µm | |
Methane sulfonamides | Methane sulfonamides | 60 Å | 3, 5, & 10 µm | |
Benzene sulfonamides | Benzene sulfonamides | 100 Å | 3, 5, & 10 µm | |
4-Nitrobenzene Sulfonamides | 4-Nitrobenzene Sulfonamides | 100 Å | 3, 5, & 10 µm | |
4-fluorobenzene sulfonamides | 4-fluorobenzene sulfonamides | 100 Å | 3, 5, & 10 µm | |
HA-Pyridines | 60 Å | 3, 5, & 10 µm | ||
HA-Dipyridyl | 100 Å | 3, 5, & 10 µm | ||
HA-DEA | 60 Å | 3, 5, & 10 µm | ||
HA-DHP | 100 Å | 3, 5, & 10 µm | ||
3,5-dihydroxyphenyl | 3,5-dihydroxyphenyl | 100 Å | 3, 5, & 10 µm | |
| ES Industries | GreenSep Ethyl Pyridine | 2-Ethylpyridine | 120 Å | 3 & 5 µm |
GreenSep Ethyl Pyridine II | A version of bonded 2-Ethylpyridine | 120 Å | 1.8, 3 & 5 µm | |
GreenSep 4-Ethyl Pyridine | 4-Ethyl Pyridine | 120 Å | 3 & 5 µm | |
GreenSep 4-Ethyl Pyridine II | A version of bonded 4-Ethylpyridine | 120 Å | 3 & 5 µm | |
GreenSep Nitro | Nitroaromatic based phase | 120 Å | 1.8, 3, 5 & 10 µm | |
GreenSep PFP | Pentafluorophenylpropyl | 120 Å | 1.8, 3, 5 & 10 µm | |
GreenSep Pyridyl Amide | Pyridyl amide | 120 Å | 1.8, 3, 5 & 10 µm | |
GreenSep Amino Phenyl | Amino + Phenyl | 120 Å | 1.8, 3, 5 & 10 µm | |
GreenSep Basic | Imidazole based phase | 120 Å | 1.8, 3, 5 & 10 µm | |
GreenSep DEAP | Diethylaminopropyl | 120 Å | 1.8, 3, 5 & 10 µm | |
GreenSep Nitro | Nitroaromatic based phase | 120 Å | 1.8, 3, 5 & 10 µm | |
GreenSep Cyano | Cyano | 120 Å | 1.8, 3 & 5 µm | |
GreenSep Diol | Diol | 120 Å | 1.8, 3 & 5 µm | |
GreenSep Naphtyl | Naphtyl | 120 Å | 1.8, 3 & 5 µm | |
GreenSep FluoroBasic | Fluorinated Imidazole | 120 Å | 3 & 5 µm | |
GreenSep HILIC | Polyhydroxylated Polymer | 120 Å | 3 & 5 µm | |
GreenSep NP-I | Optimised for the separation of 10 different cannabinoids | 120 Å | 5 µm | |
GreenSep NP-II | Optimised for the separation and isolation of THC and THCV from cannabis | 120 Å | 5 & 10 µm | |
GreenSep NP-III | Optimised for the separation and isolation of CBDA and THCA from cannabis | 120 Å | 5 & 10 µm | |
GreenSep Amine | Amino | 120 Å | 1.8 µm | |
GreenSep Silica | - | 120 Å | 1.8, 3 & 5 µm | |
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| Daicel | Chiralpak IA | Amylose tris (3,5-dimethylphenylcarbamate) | N/A | 1.6, 3, 5, & 10 µm |
Chiralpak IB | Cellulose tris (3,5-dimethylphenylcarbamate) | N/A | 1.6, 3, 5, & 10 µm | |
Chiralpak IC | Cellulose tris (3,5-dichlorophenylcarbamate) | N/A | 1.6, 3, 5, & 10 µm | |
Chiralpak ID | Amylose tris (3-chlorophenylcarbamate) | N/A | 1.6, 3, 5, & 10 µm | |
Chiralpak IE | Amylose tris (3,5-dichlorophenylcarbamate) | N/A | 3, 5, & 10 µm | |
Chiralpak IF | Amylose tris (3-chloro-4-methylphenylcarbamate) | N/A | 3, 5, & 10 µm | |
Chiralpak IG | Amylose tris (3-chloro-5-methylphenylcarbamate) | N/A | 1.6, 3, 5, & 10 µm | |
Chiralpak IH | Amylose tris (S)-α-methylbenzylcarbamate) | N/A | 1.6, 3, 5, & 10 µm | |
Chiralcel OD | Cellulose tris (3,5-dimethylphenylcarbamate) | N/A | 3, 5, & 10 µm | |
Chiralcel OJ | Cellulose tris (4-methylbenzoate) | N/A | 3, 5, & 10 µm | |
Chiralcel OX | Cellulose tris (4-chloro-3-methylphenylcarbamate) | N/A | 3, 5, & 10 µm | |
Chiralcel OZ | Cellulose tris (3-chloro-4-methylphenylcarbamate) | N/A | 3, 5, & 10 µm | |
Chiralpak AD | Amylose tris (3,5-dimethylphenylcarbamate) | N/A | 3, 5, & 10 µm | |
Chiralpak AS | Amylose tris (S)-α-methylbenzylcarbamate | N/A | 3, 5, & 10 µm | |
Chiralpak AY | Amylose tris (5-chloro-2-methylphenylcarbamate) | N/A | 3, 5, & 10 µm | |
Chiralpak AZ | Amylose tris (3-chloro-4-methylphenylcarbamate) | N/A | 3, 5, & 10 µm | |
| Sepax | SFC pyridines | Pyridines | 120 Å | 8 µm |
SFC-SCX | Sulfonic Acid & Phenyl | 120 Å | 1.8, 2.2, 3, 5, 7 & 10 µm | |
SFC-Diol | Diol | 120 Å | 1.8, 2.2, 3, 5 & 10 µm | |
SFC-Cyano | Cyanopropyl | 120 Å | 1.8, 2.2, 3, 5 & 10 µm | |
SFC-Amino | Aminopropyl | 120 Å | 3, 5, 7 & 10 µm | |
SFC-Silica | - | 120 Å | 3, 5, 7 & 10 µm | |
| Kromasil | SFC SIL | - | 100 Å | 5 & 10 µm |
SFC DIOL | Diol | 100 Å | 2.5 & 5 µm | |
SFC CN | Cyanopropyl | 100 Å | 2.5 & 5 µm | |
SFC 2-EP | 2-Ethylpyridine | 100 Å | 2.5 & 5 µm | |
SFC XT | - (Fused Organo-Silane) | 100 Å | 2.5 & 5 µm | |
| Waters | Torus 2-PIC | 2-Picalylamine | 130 Å | 1.7 & 5 µm |
Torus DEA | Diethylamine | 130 Å | 1.7 & 5 µm | |
Torus DIOL | Diol | 130 Å | 1.7 & 5 µm | |
Torus 1-AA | 1-Aminoanthracenes | 130 Å | 1.7 & 5 µm | |
Trefoil AMY1 | Amylose tris (3,5-dimethylphenylcarbamate) | N/A | 2.5 µm | |
Trefoil CEL1 | Cellulose tris (3,5-dimethylphenylcarbamate) | N/A | 2.5 µm | |
Trefoil CEL2 | Cellulose tris (3-chloro-4-methylphenylcarbamate) | N/A | 2.5 µm | |
Viridis BEH 2-EP | 2-ethylpyridine | 130 Å | 1.7, 3.5 & 5 µm | |
Viridis BEH | - (Ethylene-bridged Hybrid Particle) | 130 Å | 1.7, 3.5 & 5 µm | |
Viridis CSH Fluoro-Phenyl | Pentafluorophenylpropyl | 130 Å | 1.7, 3.5 & 5 µm | |
Viridis HSS C18 SB | C18 | 100 Å | 1.7 & 3.5 µm | |
Viridis Silica 2-EP | 2-Ethylpyridine | 100 Å | 5 µm | |
Viridis Silica | - | 100 Å | 5 µm | |
| Shimadzu | Shim-pack UC-X RP | C18 + polar Group | 100 Å | 3 & 5 µm |
Shim-pack UC-X GIS II | C18 | 100 Å | 3 & 5 µm | |
Shim-pack UC-X Phenyl | Phenyl | 100 Å | 3 & 5 µm | |
Shim-pack UC-X CN | Cyanopropyl | 100 Å | 3 & 5 µm | |
Shim-pack UC-X Diol | Diol | 100 Å | 3 & 5 µm | |
Shim-pack UC-X SiI | - | 100 Å | 3 & 5 µm | |
Shim-pack UC-X Amide | Carbamoyl | 100 Å | 3 & 5 µm | |
Shim-pack UC-X NH2 | Aminopropyl | 100 Å | 3 & 5 µm | |
| YMC | YMC-Triart Diol / | Diol | 120 Å | 1.9, 3 & 5 µm |
YMC-Triart PFP / | Pentafluorophenylpropyl | 120 Å | 1.9, 3 & 5 µm | |
YMC-Triart C18 / | C18 | 120 Å | 1.9, 3 & 5 µm | |
YMC-Triart SIL / | - | 120 Å | 3 & 5 µm | |
YMC-Pack CN / | Cyanopropyl | 120 Å | 3 & 5 µm | |
YMC-Pack SIL / | - | 120 Å | 3 & 5 µm | |
YMC-Pack 2-ethyl pyridine | 2-Ethyl pyridine | N/A | 5 µm | |
YMC-Pack Diethylaminopropyl | Diethylaminopropyl | N/A | 5 µm | |
YMC-Pack Propyl acetamide | Propyl acetamide | N/A | 5 µm | |
YMC-Pack Pyridine amide | Pyridine amide | N/A | 5 µm |
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