Supercritical trace analysis

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). 

A disadvantage of supercritical fluids for extraction is that most common fluids used for extraction (carbon dioxide, nitrous oxide, sulfur hexafluoride, etc.) are weak solvents, limiting the polarity and molecular weight range of analytes that can be efficiently extracted. Also, for trace analysis the availability of fluids of adeguate ptirity may be a problem. 

For the analysis of organic additives in polymeric materials, in most cases, prior extraction will be necessary. Depending on the nature of the additive, many different approaches are employed. These include soxhlet extraction with organic solvent or aqueous media, total sample dissolution followed by selective precipitation of the polymer leaving the additive in solution, assisted extraction using pressurised systems, ultrasonic agitation and the use of supercritical fluids. In trace analysis, solid phase extraction (SPME) from solution or solvent partition may be required to increase the analyte concentration. 

Ashraf, S., D. Bartle, A. Clifford, and R. Moulder (1991). Trace analysis of agrochemicals by supercritical fluid chromatography. J. High... 

Most FTMS instrument and method development research has been focussed on demonstration experiments. Examples include coupling FTMS with various sample introduction schemes (e.g., GC, LC, supercritical fluid chromatography), sample ionization (e.g., LD, pulsed SIMS, Cf-252 PDMS, etc.), and demonstrating application to various interesting classes of chemical compounds. These demonstrations are useful because they are indications of the potential of the technique. However, few reports of the routine use of FTMS for trace analysis, for accurate mass, and for structure determination of unknowns have yet appeared. One reason is that FT mass spectrometers are not widely spread in the hands of users. Another is that FTMS is not yet routine. Most of the demonstration experiments have been done in expert laboratories by committed and highly focussed graduate students and postdoctoral researchers. 

Table 6.13 Examples of application of supercritical fluid extraction in trace analysis...

Whilst the object of this chapter has been to show the extent and type of HPLC technique that is used today in today s environmental laboratories, there are a number of less routine techniques that may or may not have an impact on routine environmental monitoring. One of the most potentially important of these is the use of LC-MS. The problems associated with using LC-MS for trace analysis are twofold one is the usual LC-MS problem of interfacing the second is that of sensitivity of detector. The interfacing problem may well continue to have partial (compared with GC-MS interfacing) solutions such as FAB, and thermospray, etc. However, even given the advances arising from electrospray interfaces the answer may well be to move away from LC-MS to supercritical fluids and SFC-MS. 

Liu H, Wehmeyer KR. Solid-phase extraction with supercritical-fluid elution as a sample preparation technique for the ultra-trace analysis of flavone in blood plasma. J Chromatogr 1992 577 61-67. 

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. 

Greibrokk, T. and Berg, B. E. (1993) Trace analysis in capillary supercritical fluid chromatography sample introduction. Trends Anal Chem., 12, 303-8. 

The present work consists of a comparative study of three different extraction techniques—specifically, liquid-liquid extraction (LLE), solid-phase extraction (SPE), and supercritical fluid extraction (SEE)—for the trace analysis of fipronil insecticide in water samples. The extracted fipronil was analyzed via high-resolution gas chromatography using electron capture detection (HRGC-ECD). The extraction methods presented linear calibration all over the investigated concentration range (0.1-1.0 pg/L). The limit of detection (LOD) was determined at 0.1 pg/L concentration level, and precision, measured by the relative standard deviations (RSD), was 7.7% for LLE, 7.8% for SPE, and 0.5% for SEE. 

The development of methods of analysis of tria2ines and thek hydroxy metabohtes in humic soil samples with combined chromatographic and ms techniques has been described (78). A two-way approach was used for separating interfering humic substances and for performing stmctural elucidation of the herbicide traces. Humic samples were extracted by supercritical fluid extraction and analy2ed by both hplc/particle beam ms and a new ms/ms method. The new ms /ms unit was of the tandem sector field-time-of-flight/ms type. 

A method which uses supercritical fluid/solid phase extraction/supercritical fluid chromatography (SE/SPE/SEC) has been developed for the analysis of trace constituents in complex matrices (67). By using this technique, extraction and clean-up are accomplished in one step using unmodified SC CO2. This step is monitored by a photodiode-array detector which allows fractionation. Eigure 10.14 shows a schematic representation of the SE/SPE/SEC set-up. This system allowed selective retention of the sample matrices while eluting and depositing the analytes of interest in the cryogenic trap. Application to the analysis of pesticides from lipid sample matrices have been reported. In this case, the lipids were completely separated from the pesticides. 

Another application of SFC-GC was for the isolation of chrysene, a poly aromatic hydrocarbon, from a complex liquid hydrocarbon industrial sample (24). A 5 p.m octadecyl column (200 cm X 4.6 mm i.d.) was used for the preseparation, followed by GC analysis on an SE-54 column (25 m X 0.2 mm i.d., 0.33 p.m film thickness). The direct analysis of whole samples transferred from the supercritical fluid chromatograph and selective and multi-heart-cutting of a particular region as it elutes from the SFC system was demonstrated. The heart-cutting technique allows the possibility of separating a trace component from a complex mixture (Figure 12.21). 

Figure 12.21 SFC-GC heait-cut analysis of chrysene from a complex hydrocarbon mixture (a) SFC ttace (UV detection) (b) GC trace without heait-cut (100% transfer) (c) GC ti ace of heatt-cut fraction (flame-ionization detection used for GC experiments). Reprinted from Journal of High Resolution Chromatography, 10, J. M. Levy et al., On-line multidimensional supercritical fluid chromatography/capillaiy gas cluomatography , pp. 337-341, 1987, with permission from Wiley-VCFI.

Figure 12.22 SFC-GC analysis of aromatic fraction of a gasoline fuel, (a) SFC trace (b) GC ttace of the aromatic cut. SFC conditions four columns (4.6 mm i.d.) in series (silica, silver-loaded silica, cation-exchange silica, amino-silica) 50 °C 2850 psi CO2 mobile phase at 2.5 niL/min FID detection. GC conditions methyl silicone column (50 m X 0.2 mm i.d.) injector split ratio, 80 1 injector temperature, 250 °C earner gas helium temperature programmed, — 50 °C (8 min) to 320 °C at a rate of 5 °C/min FID detection. Reprinted from Journal of Liquid Chromatography, 5, P. A. Peaden and M. L. Lee, Supercritical fluid chromatography methods and principles , pp. 179-221, 1987, by courtesy of Marcel Dekker Inc.

Some typical applications in SFE of polymer/additive analysis are illustrated below. Hunt et al. [333] found that supercritical extraction of DIOP and Topanol CA from ground PVC increased with temperature up to 90 °C at 45 MPa, then levelled off, presumably as solubility became the limiting factor. The extraction of DOP and DBP plasticisers from PVC by scC02 at 52 MPa increased from 50 to 80 °C, when extraction was almost complete in 25 min [336]. At 70 °C the amount extracted increased from 79 to 95 % for pressures from 22 to 60 MPa. SFE has the potential to shorten extraction times for traces (<20ppm) of additives (DBP and DOP) in flexible PVC formulations with similar or even better extraction efficiencies compared with traditional LSE techniques [384]. Marin et al. [336] have used off-line SFE-GC to determine the detection limits for DBP and DOP in flexible PVC. The method developed was compared with Soxhlet liquid extraction. At such low additive concentrations a maximum efficiency in the extractive process and an adequate separative system are needed to avoid interferences with other components that are present at high concentrations in the PVC formulations, such as DINP. Results obtained... 

The enantiomeric purity is determined by chiral stationary phase, supercritical fluid chromatographic (CSP-SFC) analysis (Berger Instruments, Daicel Co. CHIRALCEL OD column 4% methanol, 180 psi, 3.0 mUmin flow rate detection at 220 nm). Racemic 1-phenylpropanol exhibited base-line separation of peaks of equal intensity arising from the R-isomer (tp, 2.74 min) and the S-isomer (tp, 3.10 min) whereas the synthetic alcohol showed these peaks in the ratio 97.7 / 2.3. This chromatographic method allowed for identification of the trace contaminants propiophenone (tp, 1.63 min) and benzyl alcohol (tp 3.40 min). 

Figure 8 shows a pair of typical time-resolved fluorescence decay traces for 100 / M pyrene in supercritical CO2 (Tr = 1.02 pr = 1.17). Note that the ordinate is logarithmic. The upper and lower panels show results for selective observation in the monomer (400 +. 10 nm) and excimer (460 + 10 nm) regions of the pyrene emission spectrum. Several interesting features are apparent from these traces. First, both decay processes are not single exponential. Second, the excimer emission has a significant contribution from a species that "grows in" between 30 - 75 ns this is a result of the excimer taking time to form (i.e., k in Figure 1). Third, the fits between the experimental data and the model shown in Figure 1 are good. Detailed analysis of these decay traces (10,11,21-26) yields the entire ensemble of photophysical kinetic parameters for the pyrene excimer in supercritical C02.

The NMR method we have developed gives a direct, in situ determination of the solubility and also allows us to obtain phase data on the system. In this study we have measured the solubilities of solid naphthalene in supercritical carbon dioxide along three isotherms (50.0, 55.0, and 58.5°C) near the UCEP temperature over a pressure range of 120-500 bar. We have also determined the pressure-temperature trace of the S-L-G phase line that terminates with the UCEP for the binary mixture. Finally, we have performed an analysis of our data using a quantitative theory of solubility in supercritical fluids to help establish the location of the UCEP. 

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