General sample preparation

Figure 20.1—Proportion of the time spent in each stage of a chromatographic analysis. Sample preparation generally represents a large fraction of the total time required for analysis (LC-GC Inti. 1991,4(2)).

Polypropylene HPLC sample preparation, general filtration Good for both aqueous and organic samples. Has low protein-binding properties. +++ —... 

Sample preparation Generally time-consuming Often none... 

In the case of small grains, sample preparation generally takes the form of ... 

Atomization The most important difference between a spectrophotometer for atomic absorption and one for molecular absorption is the need to convert the analyte into a free atom. The process of converting an analyte in solid, liquid, or solution form to a free gaseous atom is called atomization. In most cases the sample containing the analyte undergoes some form of sample preparation that leaves the analyte in an organic or aqueous solution. For this reason, only the introduction of solution samples is considered in this text. Two general methods of atomization are used flame atomization and electrothermal atomization. A few elements are atomized using other methods. 

Preparation of soil—sediment of water samples for herbicide analysis generally has consisted of solvent extraction of the sample, followed by cleanup of the extract through Uquid—Uquid or column chromatography, and finally, concentration through evaporation (285). This complex but necessary series of procedures is time-consuming and is responsible for the high cost of herbicide analyses. The advent of soUd-phase extraction techniques in which the sample is simultaneously cleaned up and concentrated has condensed these steps and thus gready simplified sample preparation (286). 

PL measurements are generally nondestructive, and can be obtained in just about any configuration that allows some optically transparent access within several centimeters of the sample. This makes it adaptable as an in situ measurement tool. Little sample preparation is necessary other than to eliminate any contamination that may contribute its own luminescence. The sample may be in air, vacuum, or in any transparent, nonfluorescing medium. 

Considering the numerous applications, heart-cut LC-LC has convincingly proven its value. Nevertheless, in LC-LC specific method development is generally needed for each analyte. Moreover, heart-cut procedures require accurate timing and, therefore, the performance of the first analytical column in particular should be highly stable to thus yield reproducible retention times. This often means that in LC-LC some kind of sample preparation remains necessary (see Table 11.1) in order to protect the first column from proteins and particulate matter, and to guarantee its lifetime. 

Minerals generally present difficult problems in chemical analysis, and these problems grow more serious when the elements being determined are as difficult to separate as are those named above. The time and effort that x-ray emission spectrography can save are therefore great, but there are obstacles to be surmounted. Among these are (1) Absorption and enhancement effects are often serious. (2) The element of interest may be present at low concentration in a matrix that is unknown and variable. (3) Satisfactory standards are not always easy to obtain. (4) Simple equipment sometimes does not resolve important analytical lines- completely. (5) Sample preparation and particle size often influence the intensities of analytical lines Class II deviations (7.8) can be particularly serious with minerals. 

Ceramics and minerals present many common problems, but ceramics warrant special treatment because elements of low atomic number predominate in them and they consequently offer x-ray emission spectrog-raphy of the light elements an excellent opportunity to prove its usefulness. Scott,8 in making this clear, emphasized the absorption and enhancement effects to be expected, and pointed out the need for careful sample preparation. By use of a General Electric XRD-5 spectrograph and associated equipment, he set up working curves for alumina, silica, potash, lime, phosphate, titania, and iron oxide in clays, refractories, and other ceramic materials. 

Scanning electron microscopy and replication techniques provide information concerning the outer surfaces of the sample only. Accurate electron microprobe analyses require smooth surfaces. To use these techniques profitably, it is therefore necessary to incorporate these requirements into the experimental design, since the interfaces of interest are often below the external particle boundary. To investigate the zones of interest, two general approaches to sample preparation have been used. 

The sample preparation in LC analysis is as important as the chromatographic separation itself. The procedure will often require considerable skill copied with a basic understanding of chromatographic methodology. The analyst will need to have some familiarity with micro techniques including general micro-manipulation, microfiltration, centrifugation and derivatization. 

Before the actual sample preparation procedure is described some general observations should first be made. However excellent the sample preparation and however sophisticated the equipment, the accuracy of the analysis will only be as good as the quality of the sample that is taken. If the sample is that of a reaction mixture from an organic synthesis laboratory, it is likely to be taken from a single bottle or container, by a professional chemist, and is likely to be truly representative of the bulk of the material. 

Solid samples are generally treated in one of two ways. If completely soluble, they can be dissolved directly and completely in a suitable solvent. Alternatively, if the samples contain insoluble materials that are of no interest, then they can be extracted with a selected solvent to obtain the relevant compounds in solution. The extract can be subsequently filtered or centrifuged to remove any unwanted substances that make up the sample matrix. The procedure will differ, depending on the amount of the substances present that are germane to the analysis. The preparation of samples for LC analysis from solid... 

The sample preparation is generally typical of the LC analysis of many foodstuffs although the specific substances of interest will differ widely. 

Liquid samples might appear to be easier to prepare for LC analysis than solids, particularly if the compounds of interest are present in high concentration. In some cases this may be true and the first example given below requires virtually no sample preparation whatever. The second example, however, requires more involved treatment and when analyzing protein mixtures, the procedure can become very complex indeed involving extraction, centrifugation and fractional precipitation on reversed phases. In general, however, liquid samples become more difficult to prepare when the substances are present at very low concentrations. 

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