Supercritical fluid extraction chromatography

All three types of chromatograph (gas, supercritical fluid and liquid) have been used in combination with SF extractors. The effectiveness of these combined assemblies rests on the availability of an appropriate interface between the two coupled techniques this role can be assigned to an external accumulator or to a direct connection to the chromatographic column or the retention interface. 

The function of external accumulator can be served by a six-way valve, the extract being trapped in its loop as the supercritical fluid is depressurized — the trap can optionally be packed with a sorbent. After trapping, the eluent sweeps the retained compounds to an HPLC instrument or the loop is heated to deliver the extract to a gas chromatographic column. In the latter case, the system can be used with volatile substances. 

When the extracted analytes are to be retained directly on the chromatographic column or at the retention interface, their insertion can be accomplished in various ways, namely (a) by injection into the column, whether directly (SFC, GC) or with the aid of a cooling system (GC, HPLC) (b) by split-splitless injection (SFC, GC) (c) by using a programmed temperature vaporizer (GC) or (d) by injection into a cold trap and subsequent thermal desorption (GC) or elution (HPLC). 

Although gas chromatography can only be applied to thermally stable compounds, it has been widely used in conjunction with SFE. Using these two techniques jointly, however, entails considering the following facts if proper performance is to be expected  

Both packed and capillary columns are used with the SFE-GC tandem. The detector is usually of the mass spectrometry, flame ionization or electron capture type. The most suitable interface for each application will be that allowing the extractant to be removed prior to the column — and hence to the detector. 

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B. Mumgaverl and K. J. Voorhees, On-line supercritical fluid extraction/chromatography system for trace analysis of pesticides in soybean oil and rendered fats , ]. Microcolumn Sep. 3 11-16(1991). 

Acrylonitrile metabolites have been measured in blood and urine, but, except for measurement of thiocyanate, these methods have not been developed for routine monitoring of exposed humans. Supercritical fluid extraction/chromatography and immunoassay analysis are two areas of intense current activity from which substantial advances in the determination of acrylonitrile and its metabolites in biological samples can be anticipated. The two techniques are complementary because supercritical fluid extraction is especially promising for the removal of analytes from sample material and immunoassay is very analyte-selective and sensitive (Vanderlaan et al. 1988). 

His interests have included such areas of analytical chemistry as separation science, supercritical fluid extraction, chromatography, thermal analysis, and mathematical analysis of the chemical data (chemometrics). 

Analytical Applications of Supercritical Fluid Extraction—Chromatography in the Coatings... 

Supercritical fluid extraction/chromatography can be used to characterize many of the components of todays high performance automobile coatings. For example, aliphatic isocyanates are highly reactive with protic solvents and are not chromophoric, therefore, their analysis poses a formidable task to the analytical chemist. Using carbon dioxide as the mobile phase in... 

McNally et al. have applied supercritical fluid extraction chromatography to the determination of diuron and linuron in soil [442]. Schlaeppi et al. [443] have described an automated magnetic particle-based chemiluminescent immunoassay for the determination of trisulfuron in soil [462]. 

Hagashidate, S., Yamauchi, Y., and Saito, M. (1994) Fractionation of eicosapentaenoic acid and docosahexaenoic acid esters from esterified fish oil by coupled supercritical fluid extraction-chromatography, in M. Saito, Y. Yamauchi, and T. Okuyama (eds.). Fractionation by Packed Column SFC and SFE, VCH Publishers, New York, NY, USA, pp. 191-203. 

L. T. Taylor, ACS Short Courses American Chemical Society, Supercritical Fluid Extraction/Chromatography, Blacksburg, VA, 1988. 

An example of an on-line SFE/SFC (supercritical fluid extraction/ chromatography) analysis conducted on a sample of HDPE (high density polyethylene) is shown in Figures 3 and 4. The extraction was performed with a sample size of 0.5 mg using carbon dioxide at 450 atm., at a temperature of 150°C for 30 minutes. 

Purifications of elfamycins have been described in the Hterature using Craig distribution (2,34), chromatography on Sephadex LH-20 (2,14,26) and Amberlite XAD-2 (10,17,19,26), supercritical fluid extraction (37), and chromatography on an Ito multilayer cod planet centrifuge (26,38). and nmr assignments of most elfamycins have been accompHshed (3,24,26,32). The characteristic uv spectra permits some differentiation (12) and bathochromic shifts associated with Al " complexation have been used to quantify efrotomycin (2, R = CH ) in feed premixes (39,40). 

Numerous high pressure Hquid chromatographic techniques have been reported for specific sample forms vegetable oHs (55,56), animal feeds (57,58), seta (59,60), plasma (61,62), foods (63,64), and tissues (63). Some of the methods requite a saponification step to remove fats, to release tocopherols from ceHs, and/or to free tocopherols from their esters. AH requite an extraction step to remove the tocopherols from the sample matrix. The methods include both normal and reverse-phase hplc with either uv absorbance or fluorescence detection. AppHcation of supercritical fluid (qv) chromatography has been reported for analysis of tocopherols in marine oHs (65). 

Analytical Supercritical Fluid Extraction and Chromatography Supercritical fluids, especially CO9, are used widely to extrac t a wide variety of solid and hquid matrices to obtain samples for analysis. Benefits compared with conventional Soxhlet extraction include minimization of solvent waste, faster extraction, tunabihty of solvent strength, and simple solvent removal with minimal solvent contamination in the sample. Compared with high-performance liquid chromatography, the number of theoretical stages is higher in... 

Unfortunately, not much experimental work has been carried out on the combination of Supercritical fluid extraction and liquid chromatography systems (43, 44). One of the reasons for this arises from the difficulties in achieving compatibility between the extraction solvent and the FC mobile phase. Baseline perturbations have been... 

B. Wenclawiak (Ed.), Analysis with Supercritical Fluids Extraction and Chromatography, Springer-Verlag, Berlin (1992). 

Supercritical fluid extraction (SFE) has been extensively used for the extraction of volatile components such as essential oils, flavours and aromas from plant materials on an industrial as well as an analytical scale (61). The extract thus obtained is usually analysed by GC. Off-line SFE-GC is frequently employed, but on-line SEE-GC has also been used. The direct coupling of SEE with supercritical fluid chromatography (SEC) has also been successfully caried out. Coupling SEE with SEC provides several advantages for the separation and detection of organic substances low temperatures can be used for both SEE and SEC, so they are well suited for the analysis of natural materials that contain compounds which are temperature-sensitive, such as flavours and fragrances. 

E. Ibanez, J. Palacios and G. Reglero, Analysis of tocopherols by on-line coupling supercritical fluid extraction-superaitical fluid chromatography , ]. Microcolumn Sep. 11 605-611 (1999). 

SUPERCRITICAL FLUID EXTRACTION COUPLED WITH SUPERCRITICAL FLUID CHROMATOGRAPHY... 

Y. Hirata and Y. Okamoto, Supercritical fluid extraction combined with microcolumn liquid chromatography for the analysis of polymer additives , J. Microcolumn Sep. 1 46-50(1989). 

One example of normal-phase liquid chromatography coupled to gas chromatography is the determination of alkylated, oxygenated and nitrated polycyclic aromatic compounds (PACs) in urban air particulate extracts (97). Since such extracts are very complex, LC-GC is the best possible separation technique. A quartz microfibre filter retains the particulate material and supercritical fluid extraction (SPE) with CO2 and a toluene modifier extracts the organic components from the dust particles. The final extract is then dissolved in -hexane and analysed by NPLC. The transfer at 100 p.1 min of different fractions to the GC system by an on-column interface enabled many PACs to be detected by an ion-trap detector. A flame ionization detector (PID) and a 350 p.1 loop interface was used to quantify the identified compounds. The experimental conditions employed are shown in Table 13.2. 

Although on-line sample preparation cannot be regarded as being traditional multidimensional chromatography, the principles of the latter have been employed in the development of many on-line sample preparation techniques, including supercritical fluid extraction (SFE)-GC, SPME, thermal desorption and other on-line extraction methods. As with multidimensional chromatography, the principle is to obtain a portion of the required selectivity by using an additional separation device prior to the main analytical column. 

The coupling of supercritical fluid extraction (SEE) with gas chromatography (SEE-GC) provides an excellent example of the application of multidimensional chromatography principles to a sample preparation method. In SEE, the analytical matrix is packed into an extraction vessel and a supercritical fluid, usually carbon dioxide, is passed through it. The analyte matrix may be viewed as the stationary phase, while the supercritical fluid can be viewed as the mobile phase. In order to obtain an effective extraction, the solubility of the analyte in the supercritical fluid mobile phase must be considered, along with its affinity to the matrix stationary phase. The effluent from the extraction is then collected and transferred to a gas chromatograph. In his comprehensive text, Taylor provides an excellent description of the principles and applications of SEE (44), while Pawliszyn presents a description of the supercritical fluid as the mobile phase in his development of a kinetic model for the extraction process (45). 

Figure 15.14 Separation of explosives exnacted from water by using SPE-SFE-GC at several SEE trapping temperatures, peak identification is as follows NG, nitroglycerin 2,6-DNT, 2,6-dinitrotoluene 2,4-DNT, 2,4-dinitrotoluene TNT, triniti otoluene IS, 1,3-tiichloroben-zene. Adapted Journal of High Resolution Chromatography, 16, G. C. Slack et al., Coupled solid phase extraction supercritical fluid extraction-on-line gas cliromatography of explosives from water , pp. 473-478, 1993, with permission from Wiley-VCH.

FIG. 17 Ion pair/supercritical fluid extraction (SFE) and derivatization gas chromatography-mass-spectrometry (GC-MS). 

Henry, M.C. and Yonker, C.R., Supercritical fluid chromatography, pressurized liquid extraction, and supercritical fluid extraction. Anal. Chem., 78, 390, 2006. 

High performance liquid chromatography (HPLC) has been by far the most important method for separating chlorophylls. Open column chromatography and thin layer chromatography are still used for clean-up procedures to isolate and separate carotenoids and other lipids from chlorophylls and for preparative applications, but both are losing importance for analytical purposes due to their low resolution and have been replaced by more effective techniques like solid phase, supercritical fluid extraction and counter current chromatography. The whole analysis should be as brief as possible, since each additional step is a potential source of epimers and allomers. 

Pressurized Fluid Extraction followed by Gas Chromatography/Mass Spectrometry (PFE-GC/MS) On-line Supercritical Fluid Extraction - Gas Chromatography/Mass Spectrometry (SFE-GC/MS)... 

Specific extraction methods are used to prepare the analyte for immunoassay by freeing the analyte fromboth specific and nonspecific interferences. Supercritical fluid extraction has been used to decrease the amount of solvent waste generated. Solid-phase extraction has gained popularity, and many different supports are available. One promising extraction and concentration method is immunoaffinity chromatography, which will be addressed later. 

Principles and Characteristics Supercritical fluid extraction uses the principles of traditional LSE. Recently SFE has become a much studied means of analytical sample preparation, particularly for the removal of analytes of interest from solid matrices prior to chromatography. SFE has also been evaluated for its potential for extraction of in-polymer additives. In SFE three interrelated factors, solubility, diffusion and matrix, influence recovery. For successful extraction, the solute must be sufficiently soluble in the SCF. The timescale for diffusion/transport depends on the shape and dimensions of the matrix particles. Mass transfer from the polymer surface to the SCF extractant is very fast because of the high diffusivity in SCFs and the layer of stagnant SCF around the solid particles is very thin. Therefore, the rate-limiting step in SFE is either... 

R.M. Smith and S.B. Hawthorne (eds), Supercritical Fluids in Chromatography and Extraction, Elsevier, Amsterdam (1997). 

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