Supercritical Fluid

Extraction
By
Nicole Adams
and
Morgan Campbell
Outline
 History and Background
 Theory
 Advantages
 Disadvantages
 Applications
 Conclusions
History
 First reported as high-pressure gas chromatography (HPGC) before
HPLC in 1962.
 1966 first use of supercritical CO2 as mobile phase
 Used a UV absorption detector with a quartz cell equipped with a gas-
liquid separator
 1968- used a SFC system with a mechanical backpressure regulator
that could control pressure independent of flow rate. Basic prototype
of modern packed column SFC.
 1970- development allowed pressure programming, gave a gradient.
 Overshadowed by development of HPLC in late 60’s and 70’s.
 1980’s led to commercialization of SFC instruments
◦ Open tubular columns-more like GC
◦ Packed columns-more like LC.
◦ Developed chiral separations
 1990’s- use of SFC as preparative separation
 2000’s- demands for ”green chemistry” has led to more interest in SFE
◦ Advances in column and mobile phase chemistry allowed separations of more
polar molecules

1
Supercritical Fluid

 Critical point represents the pressure and temperature conditions

under which phase such as liquid and gas cease to exist.

2
Supercritical Fluid Characteristics

 Has density and solvent power similar to

that of a liquid solvent but the viscosity
and diffusivity of the same order of
magnitude as gases

 SCF moves like a gas and dissolves

substrates similar to a liquid

2,3
Supercritical Fluid Extraction
The separation of chemicals which are mixed with a
supercritical fluid to form a mobile phase which is
subjected to pressures and temperatures near or above
the critical point for the purpose of enhancing the mobile
phase solvating power.
Typically, CO2 is used as the supercritical fluid. CO2 is
first in vapor form then compressed into a liquid prior to
becoming supercritical, where extraction occurs.

Supercritical CO2:
 Critical temperature = 30.9˚C
 Critical pressure = 73.8 bar
 Critical density = 0.467 gm/ml

2
Modifiers
 Co-solvents that are added to CO2 to enhance
extraction efficiency
 Usually 1 to 10% of methanol or ethanol is
added to expand the extraction range to
include more polar lipids
 Organic modifiers can increase the
complexity of the experimental model that
determines SCF extraction parameters

2,4
SFE Apparatus

 The necessary apparatus for a SFE setup is simple. Figure depicts the basic
elements of a SFE instrument, which is composed of a reservoir of supercritical
fluid, a pressure tuning injection unit, two pumps (to take the components in the
mobile phase in and to send them out of the extraction cell), and a collection
chamber.

5
Static Mode
 Equilibrium state between the solute and the solvent is achieved
before any sample is taken out to analyze the solubility.

 Static method carries out the equilibrium process in many ways

that include recirculation of the solvent, agitation by the
magnetic stirrer, or simply trapping the solvent in the
equilibrium cell for some time

 In the static extraction experiment, there are two distinct steps

in the process:
1. The mobile phase fills the extraction cell and interacts with the sample.
2. The second pump is opened and the extracted substances are taken
out at once.

3,5
Dynamic extraction mode

 In dynamic extraction, the second pump

sending the materials out to the collection
chamber is always open during the extraction
process.
 Thus, the mobile phase reaches the

extraction cell and extracts components in

order to take them out consistently.

5
Characteristics of SCF
 Density: the ratio of the mass of an object to
its volume
 Solubility: the maximum amount of a

substance that will dissolve in a given amount

of solvent at a given temperature to form a
stable solution
 Viscosity: the resistance of a liquid to flow
 Diffusivity: the ability of a molecule to mix

with a substance by random molecular

motion
Theory-Density
 Determines the intermolecular forces for packing of
molecules in the solvent around the molecules of solute.
This behavior determines solvation.
 Retention behavior is also related to the density of the mobile
phase, which is provided by the Equations of state (EOS)
 EOS are relationships that connect the pressure, volume, and
the temperature of a given mass of a fluid.

 Where M is the molecular weight, R the universal gas

constant, and Z the compressibility factor. Z = Z(P,T), which
is given by the EOS

3,4
Equation of State
Thereare multiple EOS equations that are very complex. Usually pressure
programs help determine what equation is the best fit for the experimental
parameters. An example is the Peng-Robinson EOS

but the volumetric mass must be calculated numerically.

Therefore,the easiest way to derive Z as a numerical solution based on this

EOS is from the equation

Where ω is the acentric factor, which can be found in tables

2, 4
Solubility
 Solvent power of a SCF depends on its structure, polarity and its density
 Solubility parameter
of a dense gas can be
estimated by:

 Where ρ/ρliq is the ratio of the density of the dense gas to that of the
liquid at its boiling point.
 Solubility increases with higher temperatures because of higher vapor
pressures but this is offset because ρ decreases with increased
temperatures and lower ρ values decrease solubility
 Initial stages of SCF extraction are governed by the distribution
coefficients of the solute between the dense fluid-phase and the
sample matrix
◦ therefore controlled by solubility

4
Viscosity
 Low viscosity enables easy penetration of the SCF in porous solids
 Viscosity of CO2 is about one order of magnitude smaller than
those of typical liquid organic solvents
 Critical viscosity can be determined by:

 Where M is molecular weight, ηc is in micropoise, Pc in atm,

Tc in K, and Vc in ml/mol
 Viscosity of a SCF essentially depends on its density which is a
function of the pressure and the temperature

4,6
Viscosity and Density
Viscosity of carbon dioxide as a function of its density at different temperatures.

 This confirms that viscosity of a fluid , subcritical, critical or supercritical depends

essentially on its density but depends little on the temperature alone.

4
Diffusivity
 SC CO2 has values that are more typical of gases
than those of the liquid state
 Increased diffusivity of an SCF as compared to
those of liquid, results in high mass transfer rates
 Self-diffusion coefficient of SC CO2 is 1-2 orders
of magnitude greater than those of dissolved
substances in the usual solvents
 Later stages of SCF extraction are governed by
diffusion-controlled process
◦ therefore controlled by mass transfer

3, 7
Diffusion Coefficients

7
Sample Matrix Influences

 Particle size and shape

 Surface area and porosity
 Moisture content
 Changes in morphology
 Sample size
 Extractable levels

2
Advantages
 “Green Chemistry”
 Can use MS, FID, UV (particularly PDA in packed columns), ESLD

 Can use gradients of CO , modifiers, density, pressure,

2
temperature
 With modifiers, can analyze a wide range of analytes
 Can be used for analytical and preparative separations
 All three parameters-pressure, temperature, modifier content-

can independently or cooperatively control retention

 CO is cheap, non-toxic, non-flammable, transmits in the UV,
2
readily available, and a gas at room temperature
 Much less use of organic solvents-good for EPA and

storage/disposal of such solvents

 CO use as a solvent protects lipid samples against oxidative
2
degradation.

1-2, 8-10
 Disadvantages
 Cost-both of equipment and training to operate machine
 Programming required to optimize results
 Equipment must be able to handle very high
pressures/temperatures
 Cannot use refractive index detection because of high back
pressure required by SFE
 Polar analytes are comparatively difficult to separate than non-
polar analytes unless a modifier is used, making the process
less “green”
 Due to temperature/pressure/”green” requirement limits, CO 2 is
the only really practical supercritical fluid solvent
 SFE is not generally selective enough to isolate specific analytes
from the matrix without further clean-up/resolution from co-
extracted species

1-2, 8-10
 Applications
 Chiral separations most successful application in SFE
(including analyte and preparative separations, has been
around longest)
 Extraction of caffeine from coffee beans, tea leaves
 Metals recovery from solids/liquids
 Food toxicology and ecotoxicology
 Solvent removal and new drug delivery formulations (used as
an anti-solvent)
 Natural pesticides
 De-nicotinization of tobacco
 Food preservatives
 Herbal medicines
 Isolation of natural products

2, 8-10
Applications (1)
 Supercritical CO2 extraction of Eucalyptus leaves
oil
 Advantages:

◦ Extracted a wide range of components-not only

volatile oils but high molecular weight compounds.
◦ 1,8-cineole (primary desired extract) content was
46.19%
◦ Extraction only took 60 minutes, compared to Soxhlet
(8 hours) or hydrodistillation (5 hours)
◦ Did not have degradation of water sensitive
compounds in oil due to partial hydrolysis
◦ No solvent residue present in finished product
8
Applications (2)
 SFECO2 extraction of bioactive compounds from
microalgae and volatile oils from aromatic plants
 Advantages:
 It is possible to obtain pure oil fraction on the second column while trapping
waxes on first column
 Extraction of bioactive compounds and volatile oils done without use of organic
solvents
 Microalga had 72g/kg hydrocarbons extracted without contamination by
chlorophyll
 These hydrocarbons can replace paraffinic and natural waxes in production of
masks for cosmetic industry-free of toxic solvents
 70% of carotenoid content extracted, used as food colorant
 Composition of extracted volatile oils from pennyroyal yielded 80% pulegone
(used as flavoring agent, in perfumery, and aromatherapy. Similar to
peppermint and camphor) and 9% menthone (related to menthol),
 Extraction of compounds from Satureja montana L. (winter savory) had 15 fold
higher amounts of thymoquinone (vs. extraction done with hydrodistillation).
Thymoquinone has anticancer, antioxidant and anti-inflammatory properties, as
well as a neuroprotective effect against Alzheimer’s disease.

9
Application (3)
 SFE of heavy metals from sand and sewage sludge
 Advantages:
◦ Lack of solvents = environmentally acceptable
◦ Morphology and structure matrix is retained
◦ Use of a chelating agent (Acetyl acetonate -dissolves in
CO2) allowed removal of Cu, Cr, Ni, Pb and Zn
◦ In comparison with traditional methods (BCR, Tessier,
etc) it is much faster
◦ Avoids analytical difficulties that are encountered with
sequential extraction methods
◦ Uses less harsh conditions than SbWE
◦ One extraction removes 30-50% metals present

10
Conclusions
 SFE is a relatively “green” process, little to no solvent in
final products
 CO can be recycled and reused
2
 In comparison to hydrodistilation and Soxhlet extraction,

SFE takes less time to extract desired compounds with less

use of organic solvents.
 Adjustments of pressure/temperature/modifiers can help

select for specific analytes

 To more fully develop SFE as a general use tool; it needs to

be cheaper, be able to be more automated, have general

lab instrumentation interface
 International Symposium on Supercritical Fluids (ISSF) held

every three years-next held in San Francisco in May 2015

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