Supercritical uids

Relatively new technique as an instrumental method. First suggested by Lovelock in 1958. First commercial instruments ! ! 1981 - packed columns 1985 - capillary columns

Supercritical uids
Two types of applications Supercritical uid extraction (SFE) Supercritical uid chromatography (SFC) As a chromatographic method, it ts somewhere between HPLC and GC. ! - offers advantages of each.

Supercritical uid advantages

As a chromatographic method can use low temperatures for separation can use packed or capillary columns pressure and polarity of mobile phase can be tuned for optimum separation As an extraction method can also be tuned removal of extracting solvent is very easy

Supercritical uid disadvantages

More expensive equipment Increased development time - more factors to control means more to optimize. Methods not as developed as GC or HPLC. ! - not as many applications (yet)

What is a supercritical uid?

Above a certain temperature, a vapor can no longer be liqueed regardless of pressure ! critical temperature - Tc
supercritical uid region

Examples
Fluid! ! Tc, oC! ! Pc, atm CO2! N2O! NH3! n-C5! n-C4! CCl2F2! ! 31.3! ! 72.9 ! 36.5!! 72.5! ! 132.5 112.5 density* ! ! 0.96 ! 0.94 ! ! 0.40 ! ! 0.51 ! ! 0.50 ! .12 1 ! -----

pressure

Pc

! 196.6! ! 33.3 ! 152.0! ! 37.5 111.8! 40.7!

solid

liquid Tc temperature gas

CHF3! ! 25.9! ! 46.9!

*density

in g/ml at 400 atm

Supercritical uids
These uids have densities and diffusivities similar to liquids but viscosities comparable to gases.
mobile phase! gas! SCF! liquid! density! (g/ml)! ~10-3! 0.2-0.9 0.8-1.0 viscosity! (poise)! 0.5 - 3.5 (x10-4) 0.2 - 1.0 (x10-3) diffusivity (cm2/sec) 0.01 - 1.0 0.1 - 3.3 (x10-4) 0.5 - 2.0 (x10-5)

Supercritical uids
This means that it is possible to develop methods that offer: ows similar to GC base methods due to low viscosity efciencies similar to HPLC methods - due to low diffusion.

0.3 - 2.4 (x10-2)

Supercritical uid extraction

With normal solvent extraction methods, solutes that have low extraction efciencies require: Batch extractions Use of large amount of solvent followed by solvent removal. Continuous extraction Heating of sample/solvent mixture. Both require heating of the solute and some venting of the solvent to the atmosphere.

Supercritical uid extraction

With SFE Can do a continuous extraction Remove solvent simply by venting Use non-toxic, inexpensive solvents Control nature of extraction by using various pressures adding modifying materials to supercritical uid

pump

extraction thimble vent

Supercritical uid extraction

Most commonly used solvent is CO2 ! ! - inexpensive - non-toxic - low critical temperature (31.3 oC)

SFC uid modier uids

collection trap restrictor nozzle

Common modiers include ! ! HCl ! make the solvent acidic. NH3! makes the solvent basic.

Supercritical uid extraction

This allows a separation to target specic types of materials

 Pressure can also be used to control extraction. Increasing pressure will increase density. Easily extracted materials will be extracted at lower densities. Raising the pressure can be used to extract materials with lower extraction efciency. The temperature of the extraction thimble can also be optimized.

Supercritical uid extraction

Example - pesticide analysis. Chromatograms obtained for two soil extracts. 2,4,5-T 2,4,5-T

extraction at 100 bar

extraction at 300 bar

Supercritical uid extraction

2,4,5-T - trichlorophenoxyacetic acid

Supercritical uid extraction

Recovery of 2,4,5-T

Supercritical uid chromatography

Supercritical uid extraction shows that we can control how a solute interacts with a mobile phase. By introducing a stationary phase, we can start doing chromatography Factors we can control - pressure of mobile phase - temperature - nature of solvent (Snyder selectivity) - stationary phase type

100 % recovery 80 60 40 20 0 50 100 150 200 250 300

Pressure can be used to alter the extraction efciency. This allows for complex samples to be extracted into groups.

Supercritical uid chromatography

150,000 capillary GC 100,000 SFC 50,000 1.0 1.1 HPLC

Supercritical uid chromatograph

programmer pressure transducer oven column detector vent restrictor

SFC ts in somewhere between GC and HPLC.

!, selectivity

1.2

pump system solvent preheater

Number of plates required to achieve a separation with RS = 1.5 and k = 2

injection valve

Supercritical uid chromatograph

Overall, equipment is similar to an HPLC. Use either a packed or capillary GC column. Either GC or HPLC type detectors can be used - based on sample and pressure. Major differences over GC and HPLC is the ability to modify pressure or solvent nature during a run.!

Pressure programming
Can be used like temperature programming for GC or gradient elution for LC. ! As pressure increases, so does the density method also called density programming. ! This results in increasing the solubility of your solutes in the mobile phase. ! So as P increases, retention decreases. Note: 1 bar = 100 kPa = 0.987 atm

Effect of pressure/density on k

Effect of pressure/density on diffusion

Diffusion Coefcient (x104)
6.000 4.725 3.450 2.175 0.900 0 0.5 1 1.5

30.0 22.5

Benzene

Benzene 55 oC

15.0 7.5 0 0.5 0.625 0.75 0.875 1

density

density

Fatty acids separated at various pressures in SC CO2

Two types of pressure programming are available. Linear - ! like temperature programming where the pressure is increased at a xed rate. Asymptotic - ! rate is decreased linearly as it approaches the maximum.

Effect of pressure

Pressure programming

Pressure Programming
pressure linear
Example. Analysis of polystyrene monomers.

Pressure gradient
150 atm/min 100 atm/min

Hydrocarbon Mixture.

pressure

asymptotic
Asymptotic programs typically result in a better separation.

Pressure gradient
Hydrocarbon Mixture.
35 atm/min

Comparison of gradients
GC, HPLC and SFC each has some sort of gradient control ! ! ! ! ! GC !- temperature

HPLC! - solvent nature/polarity SFC ! - pressure ! ! - temperature ! ! - solvent nature/polarity

SFC offers the widest range of possible control - but longest development time.

Example
Lets compare the three methods and gradients. The following are results obtained for a coal tar sample. ! ! ! ! ! ! Method! ! Gradient ! ! GC! ! ! temperature ! ! LC! ! ! polarity ! ! SFC!! ! pressure

GC - temperature gradient

LC - polarity gradient

SFC - pressure gradient

SFC modier solvents

Small amounts of a second solvent can be added to the supercritical uid. This results in a change in solvent polarity and nature - follows Snyder rules.

SFC modier solvents

Effects of polar modiers in CO2 on retention ! solute ! ! ! ! k! ACN ! ! ! CO2! IPA 1.4! ACN 1.4 CO2! IPA

proton acceptor 1 23 dipole 1 - 2-propanol 2 - acetonitrile 3 - dichloromethane

carbazole! 2.2! pyrine! ! pyrene 4-hydroxy! 9.1!

0.4! 0.2! 0.2!

4.4! 0.6! 0.3! 2.1!

1.2! ----

proton donor

0.8! ----! ----! ---- ----

7.5 mole% in CO2 at 80oC, 125 atmIPA, 135 atmACN