Diode-array

The most important applications for diode array systems are in molecular spectroscopy, since in general they do not have the resolution necessary for atomic spectroscopy. In molecular spectroscopy, the most useful areas of application are for (1) scanning fast reactions to determine kinetics, (2) applications involving low light levels because spectra can be stored and added to each other, increasing the intensity, and (3) detectors for HPLC and capillary electrophoresis (CE). HPLC and CE are discussed in Chapter 13. 

Samples for UV/VIS spectroscopy can be solids, liquids, or gases. Different types of holders have been designed for these sample types. As will be discussed in Section 5.2.6, a new class of spectrometers designed for nanoliter sample volumes does not use the standard sample cells described in the following. 

The cells or cuvettes (also spelled cuvets) used in UV absorption or mission spectroscopy must be transparent to UV radiation. The most common materials used are quartz and fused silica. Quartz and fused silica are also chemically inert to most solvents, which make them sturdy and dependable in use. (Note Solutions containing hydrofluoric acid or very strong bases, such as concentrated NaOH, should never be used in these cells. Such solutions will etch the cell surfaces, making them useless for quantitative work.) Quartz and fused silica cells are also transparent in the visible and into the NIR region, so these could be used for all work in the UV and visible regions. These are also the most expensive cells, so if only the visible portion of the spectrum is to be used, there are cheaper cell materials available, such as Pyrex . 

When double-beam instrumentation is used, two cells are needed one for the reference and one for the sample. It is normal for absorption by these cells to differ slightly. This causes a small error in the measurement of the sample absorption and can lead to analytical error. For most accurate quantitative work, optically matched cells are used. These are cells in which the absorption of each one is equal to or very nearly equal to the absorption of the other. Large numbers of cells are manufactured at one time and their respective absorptivities measured. Those with very similar absorp-tivities are designated as optically matched cells. Matched cells are usually etched near the top with an identification mark and must be kept together. It is important for the analyst to understand that even closely matched cells will show small differences in absorption due to differences in raw material characteristics. (The transmission of matched cells will also change due to normal use, so a new cell of the same match code will not necessarily match an older, used cell.) Less commonly used cells (other than the 1 cm type) can be supplied in matched sets of two or four cells. The proper use of matched cells is to fill both the sample and the reference cells with the solvent and run a baseline spectrum, which is stored by the instrument computer system. The sample cell is then cleaned and sample solution put into it, while the reference cell and its solvent are left in place. After measuring the sample spectrum, the baseline is subtracted from the sample spectrum by the computer. This approach will correct for small differences in the cells. It is also important that the sample cell be 

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Colorplate 12 shows a photo of an HPLC equipped with a diode array detector. 

In one instrument, ions produced from an atmospheric-pressure ion source can be measured. If these are molecular ions, their relative molecular mass is obtained and often their elemental compositions. Fragment ions can be produced by suitable operation of an APCI inlet to obtain a full mass spectrum for each eluting substrate. The system can be used with the effluent from an LC column or with a solution from a static solution supply. When used with an LC column, any detectors generally used with the LC instrument itself can still be included, as with a UV/visible diode array detector sited in front of the mass spectrometer inlet. 

For quantitative analysis, the resolution of the spectral analyzer must be significantly narrower than the absorption lines, which are - 0.002 nm at 400 nm for Af = 50 amu at 2500°C (eq. 4). This is unachievable with most spectrophotometers. Instead, narrow-line sources specific for each element are employed. These are usually hoUow-cathode lamps, in which a cylindrical cathode composed of (or lined with) the element of interest is bombarded with inert gas cations produced in a discharge. Atoms sputtered from the cathode are excited by coUisions in the lamp atmosphere and then decay, emitting very narrow characteristic lines. More recendy semiconductor diode arrays have been used for AAS (168) (see Semiconductors). 

Simultaneous quantification of the herbicides atra2ine, sima2ine, terbut5la2ine, propa2ine, and prometryne and their principal metabohtes has been reported in natural waters at 3—1500 ng/L concentration (104). The compounds were enriched on graphiti2ed carbon black and analy2ed with hplc and a diode array uv detector. 

The reseai ch has been carried out by the liquid chromatograph Perkin-Elmer (Series 200), which has tandem detectors the diode array (X=210 nm) and the refractometer. The temperature of a column was 30 C, speed of a mobile phase is 1.5 ml/ min. As a mobile phase, mixtures of solvents methanol - water and acetonitrile - water with addition of sodium perchlorate. The columns with the modified silica gel C8 and Cl8 (4.6x220 mm, 5 pm) were used for sepai ation of the AIST and FAS components. In order to make the identification of AIST and FAS components more reliable the ratio of the values of the above-mentioned detectors signals of each substance analyzed. 

A capillary electrophoresis systems Agilent CE 1100 (HP, USA) equipped with a diode array detector was used to separate and quantify... 

A powerful tool now employed is that of diode array detection (DAD). This function allows peaks detected by UV to be scanned, and provides a spectral profile for each suspected microcystin. Microcystins have characteristic absorption profiles in the wavelength range 200-300 nm, and these can be used as an indication of identity without the concomitant use of purified microcystin standards for all variants. A HPLC-DAD analytical method has also been devised for measurement of intracellular and extracellular microcystins in water samples containing cyanobacteria. This method involves filtration of the cyanobacteria from the water sample. The cyanobacterial cells present on the filter are extracted with methanol and analysed by HPLC. The filtered water is subjected to solid-phase clean-up using C g cartridges, before elution with methanol and then HPLC analysis. 

Separation of C oand C70 can be achieved by HPLC on a dinitroanilinopropyl (DNAP) silica (5pm pore size, 3(X)A pore diameter) column with a gradient from H-hexane to 50% CH2CI2 using a diode array detector at wavelengths 330nm (for C q) and 384nm (for C70). [J Am Chem Soc 113, 2940, 1991.]... 

Diode Array and Charge-Coupled Detector Systems... 

Figura 3 Grating spectrometers commonly used for ICP-OES (a) monochromator, in which wavelength is scanned by rotating the grating while using a singie photomultiplier tube (PMT) detector (b) polychromator, in which each photomultiplier observes emission from a different wavelength (40 or more exit slits and PMTs can be arranged along the focal plane) and (c) spectrally segmented diode-array spectrometer.

G. M. Levy, A. Quaglia, R, E. Lazure, and S. W. McGeorge. Spect. Acta. 42B, 341, 1987. Describes the diode array-based spectrally segmented spectrometer for simultaneous multielement analysis. 

The particle size analyzer, based on laser light diffraction, consists of a laser source, beam expander, collector lens, and detector (Fig. ] 3.45). The detector contains light diodes arranged to form a radial diode-array detector. The particle sample to be measured can be blown across the laser beam (dry sample), or it can be circulated via a measurement cell in a liquid suspension. In the latter case, the beam is direaed through the transparent cell. 

Photodiodes produce an electric field as a result of pn transitions. On illumination a photocurrent flows that is strictly proportional to the radiation intensity. Photodiodes are sensitive and free from inertia. They are, thus, suitable for rapid measurement [1, 59] they have, therefore, been employed for the construction of diode array detectors. 

Such effects principally cannot be observed in multi band detectors such as a UV diode array detector or a Fourier transform infrared (FTIR) detector because all wavelengths are measured under the same geometry. For all other types of detectors, in principle, it is not possible to totally remove these effects of the laminar flow. Experiments and theoretical calculations show (8) that these disturbances can only be diminished by lowering the concentration gradient per volume unit in the effluent, which means that larger column diameters are essential for multiple detection or that narrow-bore columns are unsuitable for detector combinations. Disregarding these limitations can lead to serious misinterpretations of GPC results of multiple detector measurements. Such effects are a justification for thick columns of 8-10 mm diameter. 

Figure 5.3 Analysis of 100 ml of (a) surface water and (b) drinking water sample spiked with 0.1 pig/ml of microcystins, using column-switching HPLC 1, microcystin-RR 2, microcystin-YR 3, microcystin-LR. Reprinted from Journal of Chromatography A, 848, H. S. Lee et al, On-line trace enrichment for the simultaneous determination of microcystins in aqueous samples using high performance liquid chromatography with diode-array detection , pp 179-184, copyright 1999, with permission from Elsevier Science.

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