GC Suprecritical Fluids

Theory and Instrumentation of GC
Supercritical Fluid Chromatography
Wherever you see this symbol, it is important to access the on-line course
as there is interactive material that cannot be fully shown in this reference
i manual.
Aims and Objectives
Aims and Objectives
Aims
• Introduce supercritical fluid chromatography as a powerful solution for

chromatographic analysis
• Explain fundamental aspects of supercritical fluid chromatography
• Present some fundamental hardware typically used in supercritical fluid
chromatography
Objectives
At the end of this Section you should be able to:
• Understand the benefits and limitations of supercritical fluid chromatography

• To identify some instrumentation typically used in supercritical fluid
chromatography
Content
Introduction 3
Supercritical Fluids 4
Modes of Chromatography 6
SFC Applications 7
SFC Instrumentation 12
Packed SFC 14
Capillary SFC 14
The Mobile Phase 15
Exhaust Gases 16
Organic Modifiers 17
Pumping Issues 19
SFC Columns 20
Packed Column SFC Stationary Phases 21
Capillary Column SFC Stationary Phases 22
Detection 24
Pressure Regulators 26
Advantages and Disadvantages of SFC 27
References 27
© Crawford Scientific www.chromacademy.com 2

Introduction
Supercritical fluid chromatography (SFC) is an intermediate chromatographic technique

whose properties are said to lie between gas and liquid chromatography. It utilizes
extreme conditions of temperature and pressure in such a way that the mobile phase
remains as a supercritical fluid, which has properties intermediate between a liquid and
a gas.[1]
In general selectivity terms, SFC can be regarded as a derivative of normal phase

chromatography with the added advantage of low viscosity (and high diffusivity) of
supercritical fluids, which results in high column efficiencies. Packed SFC columns are of
the same type as those currently used in normal phase chromatography including a host
of bonded phase column types which we will study later in this module.
Different eluent systems have been used to undertake SFC separations; however, carbon
dioxide based systems are by far the most successful and widely used of all of them.
When considered as a mobile phase for SFC, carbon dioxide often requires the addition of
organic modifiers (such as alcohols) for the elution of polar solutes. The term “subcritical
chromatography” denotes chromatographic separations using subcritical fluids as the
mobile phase.[2,3]
Disadvantages of SFC are mainly mobile phase or equipment related and include:[4, 5]
• Limited choice of mobile phases

• Limited analyte solubility in the mobile phase
• Unwanted reactions with the mobile phase (for example, at super-critical conditions
CO2 forms carbamic acids with primary and secondary amines)
• Repeatable and stable formation of a gradient which includes both supercritical CO2
and a polar organic modifier
HPLC separation of antipsychotics drugs.
Column: C18, 250×4.6 mm, 5 μm.

Eluent system: tetrabutyl ammonium sulphate (0.01 M) and methanol (1:1 ratio) pH 3.5,
with a flow rate of 1.5 mL/min
Detection: UV (λ = 218 nm).

SFC separation of antipsychotics drugs.
Column: C18, 250×4.6 mm, 5 μm.

Eluent system: 16.67% methanol (organic modifier) in CO2, at a flow rate of 3.0 mL/min
at a temperature of 60°C and 29.4 MPa outlet pressure
Supercritical Fluids
Any substance that is maintained above its critical conditions (heated above its critical
temperature AND compressed above its critical pressure) is said to be in a supercritical
state and substances under these conditions are termed ‘Supercritical Fluids’.[6]
Critical Pressure (Pc): is the highest pressure at which a liquid can be converted to a gas
by increasing its temperature.
Critical Temperature (Tc): is the highest temperature at which a gas can be converted to
a liquid by increasing its pressure.
Above its critical temperature a gas will not condense into a liquid phase regardless of
how much the pressure is increased; in the same way, above its critical pressure a liquid
will never exists in the gas phase regardless of how much the temperature is raised.[7]
Supercritical fluids are neither liquids nor gases; rather they are compressible fluids with
the characteristic dissolving power of liquids, but diffusivities approaching that of gases.
That is why we cannot define the supercritical fluid as a liquid or as a gas but also what
make them so interesting as a mobile phase for chromatographic separations.
Using the correct conditions of temperature and pressure and mobile phase composition
are of critical in SFC separations. However for various reasons of instrument design and
operating principle, as well as the effect of modifier addition, workers may unwittingly
operate with slightly subcritical fluids –however it is the repeatability of the applied
conditions which is important, rather the absolutely surety that one is operating
supercritically. Variations in the concentration or type of organic modifier will change the
supercritical properties of the system.

Pressure-Temperature of a pure substance (where the subscript c denotes the critical
conditions)
Critical Point: Specifies the conditions (temperature, pressure and

sometimes composition) at which a phase boundary ceases to exist.
For pure carbon dioxide the conditions of temperature and pressure that
define the critical point are: Tc = 31.1 oC, Pc = 73.8 Bar
Triple Point: The triple point of a pure substance is the temperature and
pressure at which three phases (for example, gas, liquid, and solid) of that
substance coexist in thermodynamic equilibrium.
For pure carbon dioxide the conditions of temperature and pressure that
define the triple point are: Ttr = -56.6 oC, Ptr = 5.17 Bar

Modes of Chromatography
Compared with liquids, supercritical fluids have higher diffusivity coefficients and less
resistance to mass transfer – both of which lead to sharper chromatographic peaks at
higher optimum linear velocities, which is demonstrated using the van Deemter curves
opposite.[8] As with all van Deemter plots, lower values of Reduced Plate Height indicate
higher efficiency.
Selected indicative physicochemical properties of liquids, gases and supercritical fluids

Property Liquid Gas Supercritical Fluid
Density (kg/m3) 1000 1 200-800
Viscosity (mPa s) 0.5-1.0 0.01 0.05-0.1
Diffusivity (cm2/s) 10-5 0.1 10-4 – 10-3
In general terms, the optimum linear velocity for a supercritical fluid is around three times
greater than an eluent in the condensed phase, allowing SFC separations to be carried
out more quickly without loss in efficiency (or the resulting potential loss of resolution).
Typical HPLC and SFC Van Deemter curves.
A close comparison between the physicochemical properties of supercritical fluids and

gases reveals that the former have:
• Increased densities (100 – 1000 times)

• Increased viscosities
• Stronger solvating power (as the solubility of analyte is usually related to the
mobile phase density at a given temperature)
Unlike GC, low temperatures can be used when performing SFC separations. Working at
lower temperature gives the advantage of:
• decreased retention factors

• increased potential for enantioselectivity
• permit the analysis of thermally labile samples

The preparative chromatographic separation of chiral enantiomers is one of the fastest
growing application areas in SFC.
SFC Applications
Supercritical fluid chromatography has many of the features of both liquid and gas
chromatography; therefore, it’s application area occupies an intermediate position
between the two of them.
SFC is important in situations where neither GC nor HPLC is capable of performing the
required analysis. Consider oleo-chemical compounds, they are too heavy for GC but
they usually require detection capabilities not easily found with HPLC.
The main advantage of SFC over GC comes from the adjustable nature of the elutotropic
strength of the mobile phase which can be controlled through the addition of organic
modifiers. As a consequence, SFC has extended the applicability range of
chromatography in a way never seen before.
To list the full range of SFC application areas is prohibitive, its flexibility makes it suitable
to a multitude of application types. Examples of some interesting applications are shown
below:
Analysis of paraffin:[9] Paraffin is the common name for the alkane hydrocarbons with
the general formula CnH2n+2. Paraffin wax refers to the solids with 20 ≤ n ≤ 40
SFC–AFD/FID chromatograms from an injection of 15 μg of paraffin wax dissolved in

carbon disulfide.
Column: C18, 150mm×1mm I.D, 3μm particles.

Conditions: pressure 150atm; temperature is 120◦C.
Mobile phase: 16% methanol modified (filled) SC-CO2
Detection: FID, AFD (Acoustic Flame Detection)

Food analysis:[10] Fish oil is oil derived from the tissues of oily fish. It is recommended
for a healthy diet because it contains the omega-3 fatty acids.
In-line UV SFC chromatogram of the phenacyl esters of fish oil.
Column: C18, 50mm×1mm I.D, 3.5μm particles.

Conditions: pressure 150atm; temperature is 120◦C.
Mobile phase: CO2 (phase A) and acetonitrile/Isopropyl alcohol (phase B). The modifier
profile was programmed from 1.2% B (2min) to 7.2% B at 0.3%/min.
Detection: UV

Petrochemistry:[12] Fuel oil is a fraction obtained from petroleum distillation, either as a
distillate or a residue.
SFC chromatogram diesel oil
Column: C18, 250mm×4.6mm I.D, 5.0μm particles.

Mobile phase: SFC-grade CO2.
Pressure programme: 110 atm to 160 atm for 20 min.
Detection: FID, UV
Pesticides:[13] To analyze pesticides at trace level in water samples, an extraction-

concentration step is commonly carried out before the SFC analysis.
SFC analysis of pesticides from a water sample

Mobile phase: SFC-grade CO2 plus 5% methanol
Pressure programme: 110 atm to 160 atm for 20 min.
Detection: ECD

Chiral Separations:[30] Separation of enantiomeric samples is a serious problem in
analytical chemistry; however, SFC is becoming a suitable option when dealing with this
type of samples.
Column: chiral, 250mm×4.6mm packed with the 3,5-dimethyphenylcarbamate derivative of

amylose coated on 10 μm silica-gel support.
Mobile phase: SFC-grade CO2 plus 3.0% methanol
Conditions: pressure 150 bar; temperature is 80◦C.
Detection: UV-Vis λ = 225nm
Cosmetics:[11] Beeswax is a wax formed from a mixture of several compounds. The

empirical formula for beeswax is C15H31CO2C30H61. Its main components are palmitate,
palmitoleate, hydroxypalmitate and oleate esters of long-chain (30-32 carbon atoms)
Column: C18, 15cm×4.6mm I.D, 5.0μm particles.

Mobile phase: SFC-grade CO2 (no modifier).
Detection: Flame ionization detector held at 400◦C
SFC-FID chromatogram of beeswax (0.81 mg/mL in CHCl3)

Lubricants:[14] The comprehensive analysis of the components of lubricants is a very
difficult task using current analytical tools; however, SFC is becoming important to deal
with these complex samples.
SFC/FID–UV–MS analysis of Irganox L57 (commercial car lubricant)

Mobile phase: SFC-grade CO2 plus 1.0 – 5.0% methanol
Detection: APCI(+)-MS, UV, FID

Preparative SFC:[31] Analysis of highly polar pharmaceutical products.
Preparative SFC separation of a mixture of methylphenylalanine isomers
Column: chiral, 250mm×4.6mm I.D, 5.0μm particles.

Mobile phase: 80% SFC-grade CO2 plus 20% organic modifier (1:1 methanol/ethanol)
Detection: UV-Vis λ=220nm
SFC Instrumentation
Supercritical fluid chromatography presents many of the features of both liquid and gas
chromatography; therefore, it occupies an intermediate position between the two of them.
Two major forms of SFC can be found:[17]
Capillary SFC: This mode of SFC presents similarities with conventional capillary GC.
Packed SFC: This mode of SFC presents similarities with conventional capillary HPLC.
The diagram opposite, illustrate a typical packed SFC instrument; however, from a
hardware point of view, the representation is equally valid for both capillary and packed
SFC systems.

i
Packed SFC system

Packed SFC
Packed SFC utilizes HPLC modified instrumentation to achieve supercritical separations;

nowadays, it is the most widely used of all forms of SFC. There are several key features
which distinguish Packed Column SFC instrumentation from that of a traditional HPLC
including:
• The obvious need for a tank of CO2

• Modern systems will contain equipment to regenerate the CO2 – i.e. remove the
organic modifier prior to re-circulating or re-depositing in the tank
• A back pressure restrictor placed after the analytical column
When performing packed SFC separations:
• Eluent flow rates are in the order of a few millilitres per minute
• Injection volumes range from a few to hundreds of microlitre (one aspect which
makes packed column SFC very suitable for preparative work)
A typical packed column SFC chromatographic process is depicted opposite, showing

mixing of the supercritical CO2 with an organic modifier, which is achieved in a similar way
to the mixing of different eluent components in HPLC.
Capillary SFC
Capillary column SFC shows similarity to conventional capillary GC; capillary SFC uses
GC modified instrumentation to achieve supercritical separations. When performing
capillary SFC separations:
• Eluent flow rates are in the order of a few microlitre per minute
• Injection volumes are in the order of a few hundreds of picolitre (1pL = 10-12L)
• In general, SFC columns have smaller internal diameter than their GC
counterparts
Capillary SFC system

The Mobile Phase
As we have seen, carbon dioxide is the mobile phase fluid of choice for supercritical
chromatography, the main reasons including:[15]
• Low cost
• Modest critical conditions
• Available at high purity
• Safety
• Easy to use
• Lack of another viable alternative
As well as carbon dioxide, many other fluids (like ammonia, numerous

chlorofluorocarbons and fluorocarbons, nitrous oxide, etc) have been used as mobile
phase components for SFC applications; however, up to date only carbon dioxide has
succeeded in becoming widely implemented.
The success of carbon dioxide is in great extent due to the addition of organic modifiers
(such as alcohols) that permit the analysis of polar analytes. The modifier increases the
polarity of the mobile phase to compete with the analyte for active sites on the stationary
phase, leading to reduced retention of solutes. This is highly analogous to the use of a
‘delocalising’ solvent in Normal Phase HPLC.
Critical properties of selected solvents are shown below.
Critical properties of selected solvents

Solvent Tc (oC) Pc (Bar) ρc (g/mL)
Ammonia 132.5 113.5 0.24
Benzene 289 48.9 0.3
n-Butane 152 38 0.23
Carbon dioxide 31.1 73.8 0.45
CClF3 28.8 39.5 0.58
CCl2F2 111.7 39.9 0.56
Ethane 32.2 48.9 0.2
Ethanol 243.4 63.8 0.28
Ethylene 9.3 50.4 0.22
Isopropanol 235.3 47.6 0.27
Methanol 240.5 79.9 0.27
Nitrous oxide 36.5 72.3 0.46
n-Propane 96.8 42.6 0.22
Propylene 91.9 46.2 0.23
Toluene 318.6 41.1 0.29
Water 374.2 221.2 0.34

Exhaust Gases
With the advent of preparative SFC, the consumption of carbon dioxide and organic
modifiers is constantly increasing. As expected, manufacturers are experiencing
increasing pressure to make SFC even more environmentally friendly while reducing
eluent flow rate costs by reusing it.
SFC exhaust gas collection systems usually consist of a set of decompression chambers
with temperature control. This devices permit carbon dioxide to regain its gaseous state
while the remaining components condense and are recovered. As expected, sample
components and organic modifiers tend to remain dissolved in the liquid organic modifier.
Once separated, sample components and organic modifiers can be disposed of according
to correct local disposal protocols. After purified, the flow of carbon dioxide can be
compressed and reutilised.
Note that as result of decompressing the SFC effluent, solid carbon dioxide (cardice or dry
ice) can form and clog the waste stream.
Exhaust gas collection system for SFC

Organic Modifiers
The use of modifiers will affect analyte retention and may even alter their elution order.[15]
Modifiers are added to the supercritical eluent system for the following reasons:
• To increase the polarity of the mobile phase (thus improving the solubility of polar
analytes)
• To facilitate desorption of polar analytes from the column
Analyte and organic modifier molecules will actually compete for active binding sites in the
stationary phase; therefore, analyte desorption is promoted by organic modifiers.
In practical terms only carbon dioxide with methanol or acetonitrile as modifiers are of
significant importance in SFC. However, the list of possible organic modifiers is long and
include different alcohols (with isopropanol being perhaps the most popular), cyclic ethers,
tetrahydrofuran, isopropylamine, hexane, etc.
The presence of modifiers in the mobile phase will determine its critical conditions; for
example, the addition of short chain alcohols will raise the critical temperature a few
hundred degrees Celsius (composition dependent) thus compromising the analysis of
thermally labile samples. For a given amount of substance the temperature, volume, and
pressure are interdependent variables, the relationship between them (under certain
conditions) can be represented by an equation of state.[16]
The addition of organic modifiers will preclude the use of detectors that respond to carbon
containing compounds (such as FID).
Equation of state: Equations of state can be applied to calculate

physical properties not only of pure substances but to mixtures. PVT
properties of mixtures of gases can be predicted by using equations
of state, one of them (Patel-Teja) is presented below:
RT a
P= −
V − b V (V + b) + c(V − b)
a = ∑∑ xi x j aij
i j
b = ∑∑ xi x j bij
i j
c = ∑∑ xi x j cij
i j
Where
P: pressure of the mixture
T: absolute temperature
V: Volume of the mixture
xij: mol fraction of i-component
aij, bij, cij: Patel-Teja Parameters (component dependent)
A disadvantage with using organic modifiers is that certain detection modes (such as FID),
cannot be used, due to their response to carbon containing compounds.

i
SFC separation of clenbuterol enantiomers using a chiral stationary phase.
Column: Chiral, 250×4.6 mm, 10 μm.

Eluent system: Carbon dioxide with Isopropylamine as organic modifier

Pumping Issues
In SFC the retention of a solute is influenced by the density of the mobile phase and this
in turn is highly influenced by the system pressure. Therefore the design of the pumping
system, to achieve a constant pressure, is of overriding importance in SFC.
SFC in-series dual pump design

Without any shadow of doubt, one of the most challenging aspects of SFC is accurately
pumping supercritical fluids. Nowadays, most of the available HPLC pumping systems
can handle supercritical carbon dioxide based systems; however, the actual amount of
mobile phase delivered will vary with temperature, pressure and composition (amount of
organic modifier). To overcome this problem, SFC pumping systems implement electronic
speed controlling to dynamically change the compressibility of the mobile phase.
Pumping systems lacking the ability to dynamically change compressibility will deliver
accurate flow at only one (pre-selected) pressure and composition. In applications where
the analysis conditions need to be changed (like in gradient operation), then with each
small composition change the pump will become less accurate.
Standard HPLC pumping systems usually have an inadequate compressibility

compensation range; therefore, they are unable to deliver an accurate flow of mobile
phase at supercritical conditions.
SFC Columns
SFC columns (as well as many other types of liquid chromatography columns) consist of a
stainless steel tube filled with the stationary phase. The stationary phase may be bare
silica or silica coated with a bonded phase. SFC columns use end-fittings that permit their
connection to the rest of the chromatographic system, and are identical to those used in
HPLC columns.
Packed SFC columns are usually made of stainless steel, however, ceramic columns are
also available.[32] The stationary phase is retained at each end of the tube by a sieve or
frit. SFC fittings and accessories are generally made of stainless steel.

The surface quality of the tube inside wall is of overriding importance as it affects the flow
behaviour through the column, uneven, unsmooth, or irregular surfaces will adversely
affect the separation of analytes flowing through the column.[32]
SFC capillary columns are of similar design to their GC counterparts, with the stationary
phase being chemically immobilized onto the inner wall of the capillary, which is typically
made from silica coated with a polyimide to give it strength and flexibility. These columns
have very narrow external diameters; therefore, chemical means (rendering ceramic frits)
rather than external fittings are the preferred choice to immobilise the column bed.
Packed Column SFC Stationary Phases
In general terms, SFC can be regarded as a form of normal phase chromatography with
the added advantage of low viscosity (and high diffusivity) of supercritical fluids.
Therefore, SFC columns are of the same type as those currently used in normal phase
chromatography.
For practical reasons, only carbon dioxide based systems are useful as SFC mobile
phases; therefore, SFC applications typically require polar stationary phases such as
silica amino and diol.
It is worth nothing that low to non polar stationary phases (such as octadecylsiloxane-
bonded) can also be used with SFC.
Selected commercially available SFC columns are listed below.[18]

i
Capillary Column SFC Stationary Phases
Although there are not many clear rules for capillary SFC column selection, the polarity of
the stationary phase gives an indication of its applicability and is the key parameter to
start with. However, very often different stationary phases may need to be scouted before
obtaining a proper separation, in a rather familiar way to anyone working with chiral

phases during the later 1990’s and early 2000’s. In many cases, SFC column selection
has to be performed considering previous work.[19]
Modern SFC instruments permit rapid scouting of columns of different nature while
altering (if required) the mobile phase. These systems are especially suited for column
selection under supercritical conditions and will usually contain automated switching
valves etc. One positive is the rapid short run times for SFC made possible due to the
unique properties of the eluent.
The tool opposite lists selected capillary SFC stationary phases and highlights situations
where they can be useful.

Detection
As was previously explained, SFC instruments are hybrids of their HPLC and GC
counterparts.
• Packed column SFC instrumentation is more like an HPLC system with a

reciprocating pump and a pressurized detector
• Capillary SFC systems resemble GC instruments with a syringe pump
SFC systems implement pressure restrictors (or regulators) somewhere after the column
to keep the eluent system above its critical conditions. The position of the pressure
regulator will determine the type of detection system to be used. If the pressure restrictor
is placed after the detector, then the detection system operates as in LC but at much
higher pressures – requiring the detector flow cells to be specially designed and
manufactured to withstand these pressures without cracking the cell windows or leaking.
Schematic representation of the connection of SFC and a high pressure detection system
(in this case a UV/Vis detector). Here the pressure regulator is located AFTER the
detector.

If the pressure restrictor is placed before the detector, then the detection system operates
as in GC; however, as the depressurized effluent enters the detector, noisy signals can be
rendered.
After leaving the column, the supercritical mobile phase decompresses into a gas,
allowing SFC to be coupled to many GC detection methods;[9, 21, 24, 29] however, a number
of LC devices, have also been used.[22, 23, 25, 26, 27, 28]
Schematic representation of the connection of SFC and a low pressure detection system
(in this case a FID detector). Here the pressure regulator is located BEFORE the
detector.

Pressure Regulators
Acting as pressure regulators, flow restrictors are commonly used to meet pressure
requirements of the SFC system.
The outlet of the pressure regulator is usually heated to prevent adiabatic cooling of the
expanding supercritical fluid. Adiabatic cooling of SFC mobile phases could render dry ice
(solid form of carbon dioxide) formation and flow path blocking.
A basic variable pressure regulator design includes a sensor that measures the actual
system pressure, this reading is sent to the pressure control system to be compared with
the required setting, as a result of this comparison a piston (acting as a flow restrictor) is
acted and the pressure increased or decreased as needed.
Two such devices are shown below.
SFC pressure regulator
SFC variable pressure regulator

Advantages and Disadvantages of SFC
Advantages over HPLC Disadvantages over HPLC

• Higher resolution and capacity • Limited choice of mobile phases
• Less organic solvent waste • Limited analyte solubility in the mobile phase
• Higher throughput • Unwanted reactions with the mobile phase
• Fast calibration • Unsuited for water-soluble analytes
Advantages over GC Disadvantages over GC

• Better solvation • Limited choice of mobile phases
• Extended range of samples • Unwanted reactions with the mobile phase
• The strength of the mobile phase • Hardware complexity
which can be controlled through the • The addition of organic modifiers will
addition of modifiers preclude the use of detectors that respond to
• Suitable for thermally labile samples carbon containing compounds (such as FID)
References
1. Terry A. Berger. “Separation of polar solutes by packed column supercritical fluid

chromatography” Journal of Chromatography A, 785 (1997) 3-33
2. Phyllis R. Eckard, Larry T. Taylor, Gregory C. Slack. “Method development for the
separation of phospholipids by subcritical fluid chromatography” Journal of
Chromatography A, 826 (1998) 241–247.
3. Rodger W. Stringham, Brian R. Krueger, Jonathan Marshall. “Use of elevated flow
rates in preparative subcritical fluid chromatography” Journal of Chromatography A, 1175
(2007) 112–116.
4. H. Fischer, O. Gyllenhaal, J. Vessman, K. Albert. “Reaction monitoring of aliphatic
amines in supercritical carbon dioxide by proton nuclear magnetic resonance
spectroscopy and implications for supercritical fluid chromatography” Analytical
Chemistry. 75 (2003) 62–626.
5. L.S. Daintree, A. Kordikowski, P. York. “Separation processes for organic molecules
using SCF Technologies” Advanced Drug Delivery Reviews 60 (2008) 351–372.
6. Chih Wu “Thermodynamics and Heat Powered Cycles: a Cognitive Engineering
Approach” Chapter 2. New York, Copyright © 2007 by Nova Science Publishers, Inc.
7 Miguel Herrero, Alejandro Cifuentes, Elena Ibañez. “Sub- and supercritical fluid
extraction of functional ingredients from different natural sources: Plants, food-by-
products, algae and microalgae” Food Chemistry 98 (2006) 136–148
8. Phyllis R. Brown, Eli Grushka, Susan Lunte. “Advances in Chromatography. Vol 46”
Marcel Dekker, Pp 215-220, New York 2005
9. Zhongpeng Xia, Kevin B. Thurbide. “Universal acoustic flame detection for modified
supercritial fluid chromatography” Journal of Chromatography A, 1105 (2006) 180–185
10. Isabelle François, Pat Sandra. “Comprehensive supercritical fluid
chromatography×reversed phase liquid chromatography for the analysis of the fatty acids
in fish oil” Journal of Chromatography A, 1216 (2009) 4005–4012
11. Jianjun Li. “Quantitative analysis of cosmetics waxes by using supercritical fluid
extraction (SFE)/supercritical fluid chromatography (SFC) and multivariate data analysis”
Chemometrics and Intelligent Laboratory Systems 45 (1999) 385–395
12. Akira Nomura, Joseph Yamada, and Takashi Yarita. “Supercritical-Fluid
Chromatograms of Fuel Oils on ODS-Silica Gel Column Using Fluorescence, UV-
Absorption, and Flame-Ionization Detectors” The Journal of Supercritical Fluids, 1995,8,
329-333

13 J.L. Bernal, J.J. Jiménez, J.M. Rivera, L, Toribio, M. J. del Nozal. “On-line solid-phase
extraction coupled to supercritical fluid chromatography with diode array detection for the
determination of pesticides in water” Journal of Chromatography A, 754 (1996) 145-157
14. Gwenaelle Lavison-Bompard, Didier Thiébaut, Jean-François Beziau, Bernadette
Carrazé, Pascale Valette, Xavier Duteurtre, Jean-Claude Tabet. “Hyphenation of
atmospheric pressure chemical ionisation mass spectrometry to supercritical fluid
chromatography for polar car lubricant additives analysis” Journal of Chromatography A,
1216 (2009) 837–844
15. Terry A. Berger. “Separation of polar solutes by packed column supercritical fluid
chromatography” Journal of Chromatography A, 785 (1997) 3-33
16. Chiehming J. Chang, Chany-Yih Day, Ching-Ming Ko, Kou-Lung Chiu. “Densities and
P-x-y diagrams for carbon dioxide dissolution in methanol, ethanol, and acetone mixtures”
Fluid Phase Equilibria 131 (1997) 243-258
17. Larry T. Taylor. “Supercritical fluid chromatography for the 21st century” The Journal
of Supercritical Fluids 47 (2009) 566–573
18. C.West, E. Lesellier. “A unified classification of stationary phases for packed column
supercritical fluid chromatography” Journal of Chromatography A, 1191 (2008) 21–39
19. Mohamed Maftouh, Christine Granier-Loyaux, Evelyne Chavana, Jerome Marini,
Antoine Pradines, Yvan Vander Heyden, Claudine Picard. “Screening approach for chiral
separation of pharmaceuticals Part III. Supercritical fluid chromatography for analysis and
purification in drug discovery” Journal of Chromatography A, 1088 (2005) 67–81
20 Lu Zeng, Rongda Xu, Derek B. Laskar, Daniel B. Kassel. “Parallel supercritical fluid
chromatography/mass spectrometry system for high-throughput enantioselective
optimization and separation” Journal of Chromatography A, 1169 (2007) 193–204
21. Nohora P. Vela, Joseph A. Caruso. “Element selective detection for supercritical-fluid
chromatography” Journal of Biochemical and Biophysical Methods 43 (2000) 45–58
22. Yukio Hirata, Yukinori Kawaguchi, and Yasuhiro Funada. “Refractive Index Detection
Using an Ultraviolet Detector with a Capillary Flow Cell in Preparative SFC” Journal of
Chromatographic Science, Volume 34, Number 1, January 1996, pp. 58-62
23. Susanne R. Wallenborg, Karin E. Markides, Leif Nyholm. “Oxidative and reductive
amperometric detection of phenolic and nitroaromatic compounds in packed capillary
column supercritical fluid chromatography” Journal of Chromatography A, 785 (1997)
121-128
24. Daniel G. Morgan, Kevin L. Harbol, Nicholas Peter Kitrinos, Jr. “Optimization of a
supercritical fluid chromatograph–atmospheric pressure chemical ionization mass
spectrometer interface using an ion trap and two quadrupole mass spectrometers”
Journal of Chromatography A, 800 (1998) 39–49
25. H. Shi, J.T.B. Strode III, L.T. Taylor, E.M. Fujinari. “Feasibility of supercritical fluid
chromatography-chemiluminescent nitrogen detection with open tubular columns” Journal
of Chromatography A, 734 (1996) 303-310
26. Roger M. Smith, Orapin Chienthavorn, Nicholas Danks, Ian D. Wilson. “Fluorescence
detection in packed-column supercritical fluid chromatographic separations” Journal of
Chromatography A, 798 (1998) 203–206
27. Brian J. Hoffman, Larry T. Taylor, Stephen Rumbelow, Larry Goff, J. David Pinkston.
“Separation of derivatized alcohol ethoxylates and propoxylates by low temperature
packed column supercritical fluid chromatography using ultraviolet absorbance detection”
28. Kayori Takahashi, Shinichi Kinugasa, Masaaki Senda, Koki Kimizuka, Kyoko
Fukushima, Tsutomu Matsumoto, Yasuhiro Shibata, John Christensen. “Quantitative
comparison of a corona-charged aerosol detector and an evaporative light-scattering
detector for the analysis of a synthetic polymer by supercritical fluid chromatography”

29. T. A. Berger. “Simple Correction for Variable Post Column Split Ratios using Pure
Carbon Dioxide in Packed Column Supercritical Fluid Chromatography with Independent
Pressure and Flow Control” Chromatographia 2001, 54, December (No. 11/12), 783-788
30. J.L. Bernal, L. Toribio, M.J. del Nozal, E.M. Nieto, M.I. Montequi. “Separation of
antifungal chiral drugs by SFC and HPLC: a comparative study” Journal of Biochemical
and Biophysical Methods 54 (2002) 245–254
31. Lisa M. Nogle, Charles W. Mann, William L. Watts Jr., Yingru Zhang. “Preparative
separation and identification of derivatized ß-methylphenylalanine enantiomers by chiral
SFC, HPLC and NMR for development of new peptide ligand mimetics in drug discovery”
Journal of Pharmaceutical and Biomedical Analysis 40 (2006) 901–909
32. Bernd-Walter Hoffman; Guenter Schaeufele. “Separation Column for
Chromatography” United States Patent 5651886. July 29, 1997

GC Suprecritical Fluids

Загружено:

Сведения о документе

Оригинальное название

Авторское право

Доступные форматы

Поделиться этим документом

Поделиться или встроить документ

Параметры публикации

Этот документ был вам полезен?

Это неприемлемый материал?

Авторское право:

Доступные форматы

GC Suprecritical Fluids

Загружено:

Авторское право:

Доступные форматы

Theory and Instrumentation of GC

Supercritical Fluid Chromatography

Aims and Objectives

• Introduce supercritical fluid chromatography as a powerful solution for

At the end of this Section you should be able to:

• Understand the benefits and limitations of supercritical fluid chromatography

© Crawford Scientific www.chromacademy.com 2

Supercritical fluid chromatography (SFC) is an intermediate chromatographic technique

In general selectivity terms, SFC can be regarded as a derivative of normal phase

• Limited choice of mobile phases

HPLC separation of antipsychotics drugs.

Column: C18, 250×4.6 mm, 5 μm.

© Crawford Scientific www.chromacademy.com 3

Column: C18, 250×4.6 mm, 5 μm.

© Crawford Scientific www.chromacademy.com 4

Critical Point: Specifies the conditions (temperature, pressure and

© Crawford Scientific www.chromacademy.com 5

Selected indicative physicochemical properties of liquids, gases and supercritical fluids

Typical HPLC and SFC Van Deemter curves.

A close comparison between the physicochemical properties of supercritical fluids and

• Increased densities (100 – 1000 times)

• decreased retention factors

© Crawford Scientific www.chromacademy.com 6

SFC–AFD/FID chromatograms from an injection of 15 μg of paraffin wax dissolved in

Column: C18, 150mm×1mm I.D, 3μm particles.

© Crawford Scientific www.chromacademy.com 7

In-line UV SFC chromatogram of the phenacyl esters of fish oil.

Column: C18, 50mm×1mm I.D, 3.5μm particles.

© Crawford Scientific www.chromacademy.com 8

SFC chromatogram diesel oil

Column: C18, 250mm×4.6mm I.D, 5.0μm particles.

Pesticides:[13] To analyze pesticides at trace level in water samples, an extraction-

SFC analysis of pesticides from a water sample

Column: C18, 200mm×4.6mm I.D, 5.0μm particles.

© Crawford Scientific www.chromacademy.com 9

Column: chiral, 250mm×4.6mm packed with the 3,5-dimethyphenylcarbamate derivative of

Cosmetics:[11] Beeswax is a wax formed from a mixture of several compounds. The

Column: C18, 15cm×4.6mm I.D, 5.0μm particles.

SFC-FID chromatogram of beeswax (0.81 mg/mL in CHCl3)

© Crawford Scientific www.chromacademy.com 10

SFC/FID–UV–MS analysis of Irganox L57 (commercial car lubricant)

Column: C18, 250mm×4.6mm I.D, 5.0μm particles.

© Crawford Scientific www.chromacademy.com 11

Preparative SFC separation of a mixture of methylphenylalanine isomers

Column: chiral, 250mm×4.6mm I.D, 5.0μm particles.

© Crawford Scientific www.chromacademy.com 12

Packed SFC system

© Crawford Scientific www.chromacademy.com 13

Packed SFC utilizes HPLC modified instrumentation to achieve supercritical separations;

• The obvious need for a tank of CO2

When performing packed SFC separations:

A typical packed column SFC chromatographic process is depicted opposite, showing

Capillary SFC system

© Crawford Scientific www.chromacademy.com 14

As well as carbon dioxide, many other fluids (like ammonia, numerous

Critical properties of selected solvents are shown below.

Critical properties of selected solvents

© Crawford Scientific www.chromacademy.com 15

Exhaust gas collection system for SFC

© Crawford Scientific www.chromacademy.com 16

Equation of state: Equations of state can be applied to calculate

© Crawford Scientific www.chromacademy.com 17

SFC separation of clenbuterol enantiomers using a chiral stationary phase.