The Diode Array A diode array consists of a number of photosensitive diodes place side by side and insulated from

one another in the form of a multi-layer sandwich. Each diode may be only a few thousands of an inch thick and the output from each diode can be scanned, stored and subsequently processed by a computer in a number of different ways. The common use of a diode array is to monitor light that has passed through a liquid sensor cell as in a multi-wavelength liquid chromatography detector. The light source is usually polychromatic (e.g. light from a deuterium lamp) and after passing through the cell, the light is dispersed by a quartz prism or a diffraction grating onto the surface of the diode array. Thus, each diode will receive light of a slightly different wavelength to that received by its neighbor. Those wavelengths most useful in liquid chromatography range from about 210 nm to 330 nm (i.e. UV light) and, thus, a sufficient number of diodes must be incorporated in the array to (at least) cover this range of wavelengths. Many organic compounds have characteristic spectra in the UV which can be used to help identify the substance passing though the sensor cell. Thus, when a given substance is eluted through the sensor cell, all the outputs from the array can be acquired and the result used to construct an absorption spectra that can be compared with standard spectra for identification purposes. Alternatively, by selecting the appropriate diode, the wavelength of the light at which there is maximum absorption can be selectively monitored to provide maximum detector sensitivity for that substance.

An example of the use of the variable wavelength UV in this way is afforded by the separation of some carboxylic acids that is monitored by UV absorption at 210 nm. The separation is shown in figure 29. The separation of a series of common fatty acids was carried out on a reversed phase column using water buffered with phosphoric acid as the mobile phase. Multi-wavelength dispersive detectors has proved extremely useful, providing adequate sensitivity, versatility and a linear response. As a result of the need for an optical bench inside the instrument, however, it is somewhat bulky In addition, it has mechanically operated wavelength selection and requires a stop/flow procedure to obtain spectra "on-the-fly". In contrast, the diode array detector has the same advantages but none of the disadvantages.

The Diode Array Detector

The diode array detector also utilizes a deuterium or xenon lamp that emits light over the UV spectrum range. Light from the lamp is focused by means of an achromatic lens through the sample cell and onto a holographic grating. The dispersed light from the grating is arranged to fall on a linear diode array. The resolution of the detector () will depend on the number of diodes (n) in the array, and also on the range of wavelengths covered (2 - 1). Thus Consequently, the ultimate resolving power of the diode array detector will depend on the semiconductor manufacturer and on how narrow the individual photo cells can be commercially fabricated.

The diode array detector can be used in a number of unique ways and an example of the use of a diode array detector to verify the purity of a given solute is shown in figure 31. The chromatogram monitored at 274 nm is shown in the lower part of figure 31. As a diode array detector was employed, it was possible to ratio the output from the detector at different wavelengths and plot the ratio simultaneously with the chromatogram monitored at 274 nm.

The chlorthalidone was isolated from a sample of tablets and separated by a reverse phase (C18) on a column 4.6 mm I.D., 3.3 cm long, using a solvent mixture consisting of 35% methanol, 65% aqueous acetic acid solution (water containing 1% of acetic acid). The flow rate was 2 ml/min.

A diagram of a diode array detector is shown in figure 30. Light from the broad emission source is collimated by an achromatic lens system so that the total light passes through the detector cell onto a holographic grating. In this way the sample is subjected to light of all wavelengths generated by the lamp. The dispersed light from the grating is allowed to fall onto a diode array. The array may contain many hundreds of diodes and the output from each diode is regularly sampled by a computer and stored on a hard disc. At the end of the run, the output from any diode can be selected and a chromatogram produced using the UV wavelength that was falling on that particular diode.

Figure 30. The Diode Array Detector

During chromatographic development, the output of one diode is recorded in real time producing a real time chromatogram. It is seen that by noting

the time of a particular peak, a spectrum can be obtained by recalling from memory the output of all the diodes at that particular time. If the peak was pure, the ratio of the adsorption at the two wavelengths (those selected were 225 and 245 nm) would remain constant throughout the elution of the entire peak. The upper diagram in figure 31 shows this ratio plotted on the same time scale and it is seen that a clean rectangular peak is observed whichunambiguously confirms the purity of the peak for chlorthalidone. The wavelength chosen to provide the confirming ratio will depend on the UV adsorption characteristics of the substance concerned, relative to those of the most likely impurities to be present, consequently the wavelengths must be chosen with some circumspection. Another interesting example of the use of the diode array detector to confirm the integrity of an eluted peak is afforded by the separation of the series of hydrocarbons shown in figure 32.

The separation was carried out on a column 3 cm long, 4.6 mm in diameter and packed with a C18 reversed phase on particles 3 in diameter. Courtesy of the Perkin Elmer Corporation
Figure 32. The Separation of Some Aromatic Hydrocarbons

The separation appears to be satisfactory and all the peaks appear to represent individual solutes; without further evidence, it would be reasonable to assume that all the peaks were pure. However, by plotting

the adsorption ratio, , for the anthracene peak it becomes apparent that the peak tail contains an impurity as the clean rectangular shape of the peak top is not shown. The absorption ratio peaks are shown in figure 33.

he ratio peaks depicted in figure 33 clearly indicate the presence of an impurity by the sloping top of the anthracene peak. This is further confirmed by the difference in the spectra for the leading and trailing edges of the peak.

Figure 33 Curves Relating the Adsorption Ratio,

, and Time

Spectra taken at the leading and trailing edge of the anthracene peak are shown superimposed in figure 34. Further work identified the impurity as t-butyl benzene at a level of about 5%.

Figure 35 Diode Array Spectra Demonstrating Peak Purity

A chromatogram is shown containing five peaks and spectra have been taken of peak (a) and peak (b) halfway up the rising side of each peak, at the top of each peak and halfway down the trailing edge of each peak. The spectra are also included in the figure 35. The impure and pure peak are unambiguously identified illustrating the value of this type of detecting system for analytical purposes. The performance of both types of multi-wavelength detectors are very similar and typical sensitivities would be about 1 x 10-7g/ml (significantly less than the fixed wavelength detector) with a linear dynamic range of about 5000 and a response index lying between 0.97 and 1.03.