Pre-separation

In off-line extraction the extracted analytes are collected and isolated independently from any subsequent analytical technique, which is to be employed next. For example, the extracted analyte can be collected in a solvent or on a solid sorbent. The choice of the collection method affects the possibilities for further analysis. The extracts may be used for final direct measurements (i.e. without further separation), e.g. UV and IR analysis. More usually, however, extraction is a pre-separation technique for chromatography, either off-line (the most common mode of SEE) or on-line (e.g. SFE-GC, SFE-LC-FTTR, etc.). The solvents used in extraction may affect subsequent chromatography. 

Singh, V. and Eckhoff, S.R. 1997. Economics of germ pre-separation for dry grind ethanol facilities. Cereal Chem. 74, 462 466. 

In environmental analytical applications where analyte concentrations, e.g. surfactants or their metabolites, are quite low, extraction and concentration steps become essential. Solid phase extraction (SPE) with cartridges, disks or SPME fibres (solid phase micro extraction) because of its good variety of SP materials available has become the method of choice for the analysis of surfactants in water samples in combination with FIA as well as LC—MS analysis. SPE followed by sequential selective elution provides far-reaching pre-separations if eluents with different polarities and their mixtures are applied. The compounds under these conditions are separated in the MS spectrometer by their m/z ratios providing an overview of the ionisable compounds contained in a sample. Identification in the sense it has been mentioned before, however, requires the generation of fragments. 

Here, the mixture analytical FIA-MS-MS approach reached its limitation to identify compounds. Hence, LC separations prior to MS analysis are essential to separate compounds with the same m/z ratio but with different structures. The behaviour in the LC separation will be influenced by characteristic parameters of the surfactant such as linear or strongly branched alkyl chain, the type, the number and the mixture of glycolether groups—PEG and/or PPG—and the ethoxylate chains. The retardation on SPE materials applied for extraction and/or concentration also depends on these properties and can therefore be used for an appropriate pre-separation of non-ionic surfactants in complex environmental samples as well as in industrial blends and household detergent formulations. A sequential selective elution from SPE cartridges using solvents or their mixtures can improve this preseparation and saves time in the later LC separation [22],... 

Gravity separation is little used since the equipment must be very large in order to reduce the gas velocity to a reasonably low value which allows the finer particles to settle. For example, a settling chamber designed to remove particles with a diameter of 20. im and density 2000 kg/m3 from a gas stream flowing at 10 m3/s would have a volume of about 3000 m3. Clearly, this very large volume imposes a severe limitation and this type of equipment is normally restricted to small plants as a pre-separator which reduces the load on a more efficient secondary collector. 

The low flow rate in the microbore column ensures sample volumes compatible with the secondary conventional column and permits the injection of a small volume onto the secondary column, making the transfer of incompatible solvents possible without peak shape deterioration or resolution losses [63], The possible disadvantage could be the lower sample capacity of microbore LC columns. However, in LCxLC, a sensitivity enhancement can be obtained if the formation of compressed solute bands at the head of the secondary column is achieved during the transfer from the first to the second dimension. Moreover, a larger volume can be injected into the first-dimension microcolumn, used as a highly efficient pre-separation step, and a limited decrease in efficiency due to a large injection volume can be tolerated. 

Some of the above mentioned problems can be reduced by applying pretreated samples to the column. The samples may be partially digested or passed through a precolumn It is therefore essential to provide information showing that the metal-ligand interaction is not disrupted during the pre-separation steps and no change in the state of the metal has occured. 

The ideal application for MIPs appears to be in SPE. SPE is generally used to improve the selectivity of analytical measurements by pre-separating the analytes from many other matrix components. In some cases the analytes are also preconcentrated on the SPE column. 

This approach has been used extensively for amino acid analysis using low-pressure ion-exchange chromatography and post-column ninhydrin reaction. Spraying, dipping and vapour-treatment techniques are well known as post-separation reactions in TLC, but these are considered only briefly since the majority of them are not quantitative. While the problems of pre-separation techniques are quite similar for TLC and HPLC, they differ considerably for post-separation reactions. 

---