Supercritical fluid extraction quantitative analytical

Hawthorne, S.B. Miller, D.J. Burford, M.D. Langenfeld, J.J. Eckert-Tilotta, S. Louie, P.K. Factors controlling quantitative supercritical fluid extraction of analytical samples. J. Chromatogr., A 1993, 642 (1-2), 301-317. 

Supercritical fluid extraction (SFE) and Solid Phase Extraction (SPE) are excellent alternatives to traditional extraction methods, with both being used independently for clean-up and/or analyte concentration prior to chromatographic analysis. While SFE has been demonstrated to be an excellent method for extracting organic compounds from solid matrices such as soil and food (36, 37), SPE has been mainly used for diluted liquid samples such as water, biological fluids and samples obtained after-liquid-liquid extraction on solid matrices (38, 39). The coupling of these two techniques (SPE-SFE) turns out to be an interesting method for the quantitative transfer... 

Supercritical fluids were soon found to be highly efficient extraction media, chiefly because of their high solvating power, their low viscosities (intermediate between a gas and a liquid), and their low surface tensions that enable their penetration deep into the extraction matrix. Supercritical fluid extraction (SEE) used in isolation is generally not selective enough to separate specific solutes from the matrix without further cleanup or resolution from coextracted species prior to qualitative and quantitative analysis. Consequently, for analytical applications, SFE is usually used in combination with chromatographic techniques to improve the overall selectivity in the isolation of specific solutes. The combined use of SFE with chromatographic techniques is quite recent. 

Finally, supercritical fluid chromatography, in which a supercritical fluid is used as the mobile phase, was introduced by Klesper [164-166]. SFE directly coupled to SFC provides an extremely powerful analytical tool. The efficient, fast and selective extraction capabilities of supercritical fluids allows quantitative extraction and direct transfer of the selected solutes of interest to be accomplished to the column, often without the need for further sample treatment or cleanup. Extraction selectivity is usually achieved by adjusting the pressure of the supercritical fluid at constant temperature or, less often, by changing the temperature of the supercritical fluid at constant pressure. SFE coupled with packed column SFC has found... 

Chlorinated phenolic compounds in air-dried sediments collected downstream of chlorine-bleaching mills were treated with acetic anhydride in the presence of triethylamine. The acetylated derivatives were removed from the matrix by supercritical fluid extraction (SEE) using carbon dioxide. The best overall recovery for the phenolics was obtained at 110°C and 37 MPa pressure. Two SEE steps had to be carried out on the same sample for quantitative recovery of the phenolics in weathered sediments. The SEE unit was coupled downstream with a GC for end analysis . Off-line SEE followed by capillary GC was applied in the determination of phenol in polymeric matrices . The sonication method recommended by EPA for extraction of pollutants from soil is inferior to both MAP and SEE techniques in the case of phenol, o-cresol, m-cresol and p-cresol spiked on soil containing various proportions of activated charcoal. MAP afforded the highest recoveries (>80%), except for o-cresol in a soil containing more than 5% of activated carbon. The SEE method was inefficient for the four phenols tested however, in situ derivatization of the analytes significantly improved the performance . 

Off-line supercritical fluid extraction, ultrasonic supercritical fluid extraction, and on-line supercritical fluid extraction-gas chromatography methodologies that have been developed specifically for analytical sample preparation and analysis are described. These methods offer the potential for extraction rate increases of over an order of magnitude, and are compatible with online analysis which provides the basis for automated sample preparation and analysis. These methods are particularly useful for small sample sizes or trace levels of analytes, and have been demonstrated to operate quantitatively. Combined ultrasonic supercritical fluid extraction can further enhance extraction rates from macro-porous materials by inducing convection through internal pores. The apparatus and instrumentation are described in detail and several examples are presented illustrating the applicability of these methodologies. 

In this context, studies about the development of relevant analytical methods allowing the detection of pesticide residues in VOO are usually focused on an optimization of the various steps of the analysis process, namely extraction, clean-up, identification, and quantitation of pesticide content. The common extraction methods are Soxhlet extraction, microwave-assisted extraction (MAE), supercritical fluid extraction (SEE), and accelerated solvent extraction (ASE). Cleanup methods include SPE, matrix solid-phase dispersion (MSPD), and gel permeation chromatography (GPC). 

Metabolites of chlorobenzene in biological materials cannot be determined in routine practice because of the lack of standard methods for measuring these metabolites. Further research on supercritical fluid (SCF) extraction holds great promise for meeting the goals of quantitative, rapid, easily performed, low cost, and safe procedures for the determination of nonpolar organic analytes such as chlorobenzene in biological samples. 

In closing, this paper was not intended to represent an exhaustive process development effort in flavors extraction from natural materials nor a development of the quantitative analytical capabilities of supercritical carbon dioxide. However, even though the examples and the conditions of extraction were somewhat arbitrary, they point out some of the interesting features of the pressure dependent dissolving power properties of supercritical fluids. They can be further refined by virtue of more narrow ranges and ratios of pressure and temperature to accomplish still more narrow separations. 

Increased amounts of sample call for proportionally increased volumes of supercritical fluid. The sample volume and extraction time can have strong effects in those cases where the analyte concentration in the sample is quite high. Figure 7.8A illustrates the recovery of PCBs from river sediment achieved with pure supercritical CO,. As can be seen, recovery of the target analytes was almost quantitative after 50 min of dynamic extraction of 100 mg of sample on the other hand, quantitative extraction from 1 g of sample was impossible even after 120 min, possibly because the extractant was saturated with PCBs [38]. 

When the analytes are to be retained in a sorbent, the sample (which can be solid, semi-solid, liquid or gaseous) is inserted in the solid state into the extraction cell. Samples in the latter three forms are supported on an appropriate material in order to ensure effective attack by the supercritical fluid. Solid supports are not used for liquid, gaseous and semi-solid samples only, however. Some research work conducted so far on solid samples has involved not natural samples but synthetic ones prepared from a selected sorbent (a natural matrix where the presence of the analytes of interest was previously excluded or a synthetic support such as polyurethane foam or glass wool) with which a solution containing the analytes was homogenized. Quantitative evaporation of the analyte solvent is mandatory as any residual solvent may alter the polarity of the supercritical fluid and hence its action to an extent dependent on the particular fluid and solvent properties, and also on the amount of solvent retained. 

The choice of the most suitable instrumental technique depends on several factors, such as the physical-chemical characteristics of analytes, the detection limits required, the level and type of interferences, the resolution needed, the identification power required, the accuracy and the precision of the quantitative determination, the availability of instrumentation and finally the cost and the time necessary per each determination. Moreover, extraction and clean-up procedures have to be suitably matched with instrumental analysis. GC coupled with Electron Capture Detection (ECD) or Mass Spectrometry (MS) has been widely applied for the determination of PCBs in organic extracts of environmental samples. In few cases the instrumentation includes the extraction step, such as an SEE system coupled with Supercritical Fluid Chromatography (SFC) or with GC (40). 

With fluids, we think of the pump as the source of pressure as well as the flow rate determining device. However, with supercritical fluids (in contrast to t3q)ical liquids), a pump needs a control point downstream to hmit the passage of molecules per unit time. This restriction then "holds-back" the previously unlimited flow of molecules to a definite, but not always pre-determined level. Ideally then, the restrictor serves to restrict the flow until the density of molecules distributed from the pump through the extraction region right up to the final restriction point in space is such that the operating density desired in the extraction zone is achieved. This is much easier to state in words than it is to achieve in actual experimental practice. This is especially true if you wish to achieve an experimental set of parameters and hold those values over a finite period of time (ranging from minutes to hours) and do it with the statistical precision and accuracy that are necessary to attain the final quantitative analytical results. 

The most common extraction techniques for semivolatile and nonvolatile compounds from solid samples that can be coupled on-line with chromatography are liquid-solid extractions enhanced by microwaves, ultrasound sonication or with elevated temperature and pressures, and extraction with supercritical fluid. Elevated temperatures and the associated high mass-transfer rates are often essential when the goal is quantitative and reproducible extraction. In the case of volatile compounds, the sample pretreatment is typically easier, and solvent-free extraction methods, such as head-space extraction and thermal desorption/extraction cmi be applied. In on-line systems, the extraction can be performed in either static or dynamic mode, as long as the extraction system allows the on-line transfer of the extract to the chromatographic system. Most applications utilize dynamic extraction. However, dynamic extraction is advantageous in many respects, since the analytes are removed as soon as they are transferred from the sample to the extractant (solvent, fluid or gas) and the sample is continuously exposed to fresh solvent favouring further transfer of analytes from the sample matrix to the solvent. 

When on-column injection is used the end of the transfer capillary is inserted into the column inlet or retention gap where decompression of the supercritical fluid occurs. Carbon dioxide gas exits through the column and the seal made between the restrictor and septum (unless a closed injector is used). The analytes are focused by cold trapping in the stationary phase. The transfer line must be physically removed from the injector at the completion of the extraction to establish the normal carrier gas flow for the separation. Analyte transfer to the column is virtually quantitative but blockage of the restrictor is more conunon and involatile material accumulates in the injection zone eventually degrading chromatographic performance. The on-column interface is probably a better choice for trace analysis of relatively clean extracts with modest fluid flow rates than the split interface. When optimized both the on-column and split interfaces provide essentially identical peak shapes to those obtained using conventional solution injection. 

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