On-line detection, use

Mincsovics and Tyihak summarized (64) the possible combinations of off-line and on-line OPLC. The fastest separations can be obtained in the fully on-line mode (on-line sample application and on-line detection) using chromatoplates with a concentrating zone. This operating mode is also the simplest and most economic method of preparative OPLC since, after cleaning and reequilibration, the same plate may be used several times without loss of resolution. Zogg et al. (65) showed that, on the basis of the experience gained in the HPLC separations, fully on-line OPLC separations were carried out with the same mobile phase composition. The results show that practically the same resolution and peak order as in HPLC could be achieved, but in a longer separation time. 

Figure 8.19 illustrates another example of the versatility of multidimensional OPLC, namely the use of different stationary phases and multiple development ("D) modes in combination with circular and anticircular development and both off-line and on-line detection (37). Two different stationary phases are used in this configuration. The lower plate is square (e.g. 20 cm X 20 cm), while the upper plate (grey in Figure 8.19) is circular with a diameter of, e.g. 10 cm. The sample must be applied on-line to the middle of the upper plate. In the OPLC chamber the plates are covered with a Teflon sheet and pressed together under an overpressure of 5 MPa. As the mobile phase transporting a particular compound reaches the edge of the first plate it must-because of the forced-flow technique-flow over to the second (lower) stationary phase, which is of lower polarity. 

LC-MS with on-line SPE using a RAM pre-column with an internal ODS phase was described by van der Hoeven et al. (95) for the analysis of cortisol and prednisolone in plasma, and arachidonic acid in urine. The samples were injected directly and the only off-line pretreatment required was centrifugation. By using the on-line SPE-LC-MS system, cortisol and related compounds could be totally recovered and quantified in 100 p.1 plasma within 5 min with a typical detection of 2 ng/ml (Figure 11.6(b)). The RAM-type of sorbents, in which the outer surface of the particles is covered with aj-acid glycoprotein, also appear to be useful for direct SPE of... 

Smith and Udseth [154] first described SFE-MS in 1983. Direct fluid injection (DFT) mass spectrometry (DFT-MS, DFI-MS/MS) utilises supercritical fluids for solvation and transfer of materials to a mass-spectrometer chemical ionisation (Cl) source. Extraction with scC02 is compatible with a variety of Cl reagents, which allow a sensitive and selective means for ionising the solute classes of interest. If the interfering effects of the sample matrix cannot be overcome by selective ionisation, techniques based on tandem mass spectrometry can be used [7]. In these cases, a cheaper and more attractive alternative is often to perform some form of chromatography between extraction and detection. In SFE-MS, on-line fractionation using pressure can be used to control SCF solubility to a limited extent. The main features of on-line SFE-MS are summarised in Table 7.20. It appears that the direct introduction into a mass spectrometer of analytes dissolved in supercritical fluids without on-line chromatography has not actively been pursued. 

For all CE instruments that use on-line detection at a fixed point along the capillary, CIEF must include a means of transporting the focused zones past the... 

Ti, or PEEK (polyether ether ketone) to allow measurements under very corrosive conditions. The separated phases pass AMX gadgets for on-line detection (radiometric, spectrophotometric, etc.) or phase sampling for external measurements (atomic absorption, spectrometric, etc.), depending on the system studied. The aqueous phase is also provided with cells for pH measurement, redox control (e.g., by reduction cells using platinum black and hydrogen, metal ion determination, etc.) and temperature control (thermocouples). 

On-line anion exchange LC/ICP-MS methods for selenium and chromium speciation have been published [30], In studies of selenium speciation in environmental samples LC/ICP-MS and LC/ ESI-MS methods have been used for the determination of both inorganic and organic selenium species [43]. For the determination of methyl selenide, strong anion exchange and RP chromatography have been employed [43], while for the determination of seleno-aminoacids, IP RP chromatography with on-line detection based on ICP-MS has been successfully employed [44],... 

CE detection is similar to detectors in, and include absorbance, fluorescence, electrochemical, and mass spectrometric detectors. The capillary can also be filled with a gel, which eliminates the electroosmotic flow. Separation is accomplished as in conventional gel electrophoresis but the capillary allows higher resolution, greater sensitivity, and on-line detection. In CE, low picogram amounts of analytes can be detected using glass fiber optics. However, this does not mean low limits of detection since only a few nanoliters can be injected. 

In the future, one can anticipate increases in sensitivity, specificity and laboratoiy-to-laboratory reproducibility of cost effective and user friendly IA s. Growth will be witnessed in the areas of (i) immunoaffinity chromatography to selectively isolate, enrich or remove components from foods, (ii) immunocytochemistry to localize and understand role of ingredients in foods, (iii) process control with advent of sensors and other on-line detection systems, and (iv) alternate uses. 

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