Supercritical fluid chromatographic method

Chromatography. GC is the most common anal)d ical method used but liquid and supercritical fluid chromatographic methods are being increasingly developed. Like titration the sample is destroyed in the analysis process. The ideal situation depicted in Figure 8.8 cannot normally be applied for titration or chromatographic analysis since the analysis equipment needs to be close to the sampling device. This is often termed at-line analysis. 

Following their extraction and cleanup, residues of sulfonamides and diaminopyrimidine potentiators in sample extracts can be detected by direct nonchro-matographic methods, or thin-layer, gas, liquid, or supercritical fluid chromatographic methods (Table 29.7). 

General - A comprehensive, critical review (228 refs.) has covered developments over the last ten years in the chromatographic analysis of carbohydrates by g.c., h.p.l.c., t.l.c., and supercritical fluid chromatographic methods. T.l.c., h.p.l.c., and g.c.-m.s. methods for analysis of cardiac glycosides, and h.p.l.c. and g.c. methods for the analysis of nucleoside antiviral drugs, have also been reviewed. 

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). 

Berger, T.A. Fogleman, K. Staats, T Bente, P Crocket, L Farrell, W. Osonubi, M. The Development of a Semi-preparatory Scale Supercritical-fluid Chromatograph for High-throughput Purification of Combi-Chem Libraries, J. Biochem. Biophys. Methods 43, 87-111 (2000). 

Berger TA, Fogelman K, Staats T, Bente P, Crocket I, Farrel W, Osonubi M. The development of a semi-preparatory scale supercritical-fluid chromatograph for high throughput purification of combi-chem libraries. J Biochem Biophys Methods 2000 43 87-111. 

In the early years, the petroleum industry acted as a catalyst for the development of this physical method. However, over the past twenty-five years, organic mass spectrometry has been the subject of a series of major developments. Some of them, such as the advent of commercially available interfaces for gas-liquid, high performance liquid, and supercritical fluid chromatographs, as well as novel ionisation techniques particularly well suited for high molecular weight, non-volatile macromolecules were welcome and well-accepted by the food analysts. Others, such as the introduction of relatively inexpensive quadrupole mass filters that proved to be reliable and... 

A very recent volume edited by Berthed (2002) is on countercurrent chromatography - the support-free liquid stationary phase. Ebdon et al. (1987) review directly coupled liquid chromatogramphy-atomic spectroscopy. The review by Uden (1995) on element-specific chromatographic detection by atomic absorption, plasma atomic emission and plasma mass spectrometry covers the principles and applications of contemporary methods of element selective chromatographic detection utilizing AA, AES and MS. Flame and furnace are considered for GC and HPLC, while MIP emission is considered for GC and ICPAES for HPLC. Combinations of GC and HPLC with both MIPAES and ICPAES are covered and supercritical fluid chromatographic (SFC) and field flow fractionation (FFF) are also considered. 

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. 

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). 

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... 

MULTIDIMENSIONAL CHROMATOGRAPHIC METHODS WHICH INVOLVE THE USE OF SUPERCRITICAL FLUIDS... 

In order to reduce or eliminate off-line sample preparation, multidimensional chromatographic techniques have been employed in these difficult analyses. LC-GC has been employed in numerous applications that involve the analysis of poisonous compounds or metabolites from biological matrices such as fats and tissues, while GC-GC has been employed for complex samples, such as arson propellants and for samples in which special selectivity, such as chiral recognition, is required. Other techniques include on-line sample preparation methods, such as supercritical fluid extraction (SFE)-GC and LC-GC-GC. In many of these applications, the chromatographic method is coupled to mass spectrometry or another spectrometiic detector for final confirmation of the analyte identity, as required by many courts of law. 

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). 

Scheme 7.3 Generalised supercritical fluid methods for extraction and analysis with and without separation of interfering components. After King [5], Reproduced from the Journal of Chromatographic Science, by permission of Preston Publications, a Division of Preston Industries, Inc.

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