Instruments, detection levels combined

The ultimate in selectivity in HPLC detection is seen with the use of mass-spectrometric detection, and for many applications this could be seen as the ideal detection method. However, more mundane considerations such as size of the instrumentation and limited budgets combine to reduce HPLC-MS to a relatively small number of applications which most effectively exploit its unique properties. When such practical constraints are taken into account, the real detector coimected to the HPLC system usually turns out to be a device that is a compromise, and its performance characteristics need to be taken into account during the development of many analyses just as much as the performance of the column or any other component of the HPLC system. For example, lack of detection selectivity may require extra method development to completely resolve an interfering peak, or lack of sensitivity could force the inclusion of an extraction-concentration step in an analytical method to achieve detectable levels of analyte. 

LOQ) will typically be higher than the instrumental detection limit (IDL), because of background analyte and matrix-based interferences. The BEC (blank equivalent concentration) used in Table 4.7 is the apparent concentration of an analyte normally derived from intercepted point of its calibration curve or by reference of the actual counts for that analyte in a blank solution. The BEC gives a good indication of the blank level, which will affect the IDL. Most often, the detection limits are calculated as three times the normal standard deviation of the BEC in a within batch replicate analytical measurement of a blank solution. Therefore, if the instrument is stable enough, this will give a better IDL than the BEC itself. The BEC is a combination of the contamination of the analyte in the solution, the residual amount of the analyte in the spectrometer and the contribution of any polyatomic species in the analyte mass. 

Selectivity and sensitivity of available instruments are tested in all laboratories in the initial step of validation. The crops used for fortification experiments and the concentration levels are identical in all laboratories. Recoveries are determined with all available detection techniques, but after discussion of the results each laboratory selects individually one valid result for each analyte-matrix-level combination. Only this result is used for the calculation of the final mean recovery and standard deviation. Typical criteria for the acceptance of methods are given in Table 11. 

The versatility and the robustness of CE separation in conjunction with the extreme sensitivity inherent to CL-based reactions make a combination of both techniques promising for application in a wide range of fields, including environmental analysis, biomedicine, and biological research and practice. Obviously, in comparison with other detection modes widely incorporated in CE, CL detection is a slowly evolving technique and advances should focus on the development of new detectors that are instrumentally simpler than existing systems and that offer the ability to detect various types of analytes at trace levels. 

Experience at Barringer Research Laboratory demonstrated that the most effective method to reduce stray light is to combine several reduction procedures. The lead/calcium selectivity of the instrumentation as received was 160, but this has been increased to greater than 1,000,000 with the manufacturer s modifications. The actual steps included replacement of the calcium 393.3 nm line with the 315.9 nm line, replacement of the lead 405.8 nm line with the 220.3 nm line, installation of an interference filter mask over the lead photomultiplier, and computer correction of the residual calcium interference. The stray light reduction obtained by installation of interference filters is presented for three common concomitants in Table II. In many cases the stray light levels were less than or equivalent to the detection limit. The interference filters and the photomultiplier masks (which reduce the entrance angle to the photomultiplier to include only the receiver mirror) improved the detection limits for many elements in the array by decreasing the system band pass. 

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