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Introduction
SFC is a hybrid of gas and liquid chromatography that combines some of the best features of both.
In SFC, the sample is carried through a separating column by a supercritical fluid where the mixture is divided based on the interaction between analytes and a stationary phase in the column.
SF has densities, viscosities and other properties that are intermediates between those of the substance in its gaseous and liquid state
SF:
- diffusivity much higher than a liquid readily penetrates porous and fibrous solids - Low viscosity (equal to gas)
Inexpensive, innocuous and non-toxic Higher diffusion coefficients and lower viscosities (compared to liquid)
faster and higher resolution separations
Components of SFC
SFC Columns
Two types of analytical columns used in SFC: (1) Capillary columns of fused silica coated with cross-linked chemically bonded stationary phases, that are used in GC are equally applicable in SFC.
(2) Packed columns developed for high performance liquid chromatography (HPLC) are being used with SFC.
(2) Packed columns developed for high performance liquid chromatography (HPLC) are being used with SFC. (a) Silica gel, the major material for current phases
Irregularly-shaped particles
Organochlorosilane
- If R is a polar functional group, the stationary phase will be polar. i.e. cyano (C2H4CN), diol (C3H6OCH2CHOHCH2OH), or amino (C3H6NH2)
- If R is a non polar functional group, the stationary phase will be non polar. i.e. n-octyl (C8) or n-octyldecyl (C18) hydrocarbon chain
Aluminium oxide Al2O3 Zirconium oxide ZrO2 Styrene-divinylbenzene Hydroxymethylstyrene Porous graphite
SFC Injection
-For packed SFC, a typical LC injection valve is commonly used
load
Inject
- In capillary SFC, small sample volumes must be quickly injected into the column and therefore pneumatically driven valves are used.
Loading Injection
Column
Sample in
Waste/Drain
SFC Pump
The type of high pressure pump used in SFC is determined by the column type. - For packed columns, reciprocating pumps are generally used
Reciprocating pumps allow easier mixing of the mobile phase or introduction of modifier fluids.
Stroke length Plunger diameter Stroke volume Pumping mechanism dual head reciprocation
SFC Detector
FID (flame ionization detector) and MS detector. The restrictor is installed before the detector, UV-absorption, fluorescence or light diffusion. The restrictor is installed after the detector.
Restrictor: a device installed in the SFC system to control pressure of the fluid during the chromatographic process
SFC Vs LC Instrumentations
The major difference in SFC and conventional LC equipment is the pumping systems as well as the safety features installed to maintain higher pressure. Unique SFC equipment differences are:
1. Carbon dioxide tank for mobile phase supply - Equipped with a pressure relief value and rupture disk
2. High-pressure pump - Chiller to maintain mobile phase in liquid state 3. High-speed injector 4. Pressure restrictor - High-pressure tubing 5. High-pressure flow cell for UV detection 6. Solvent collection device with ability to vent to a laboratory hood or elephant trunk.
and contain no functional group that makes possible detection in LC using spectroscopic or electrochemical techniques < up to 25% of all separation problems fall into this category < examples include: polymers, fossil fuels, pesticides, foods, drugs, etc - Separations are faster then LC - Run at lower temperature than GC - Beneficial in industrial scale purification
Faster elution
Effects of Pressure:
Pressure increases results in reduced elution time - increase in density of mobile phase - effects retention or capacity factor (k) - pressure changes analogous to gradient elution in LC and GC
Application of SFC
SFC is more and more used in research and development laboratories and pilot plants of the pharmaceutical and fine chemical industries. SFC is particulary interesting for the purification of : chiral compounds actives or intermediates from complex mixtures lipophilic compounds The ability to vary selectivity by programming the parameters P (pressure) and T (temperature) rather than by modifying the chemical composition of the eluent. The range of compounds analysed by SFC includes lipids and oils, emulsifiers, oligomers and polymers (compounds of molecular mass greater than 1000 which cannot be studied in GC)
(1) Chiral separation The use of supercritical fluids to separate enantiomers is one of the most important tasks in several areas of research, especially pharmaceuticals and agrochemicals. It is well known that the two enantiomeric forms of a molecule can display dramatically different biological activity.
- Showed the effects of different organic modifiers on the resolution and retention via k value