As already mentioned, a special feature of some variable wavelength UV detectors is the ability to perform spectroscopic scanning and precise absorbance readings at a variety of wavelengths while the peak is passing though the flowcell. Diode array adds a new dimension of analytical capability to liquid chromatography because it permits qualitative information to be obtained beyond simple identification by retention time.
There are two major advantages of diode array detection. In the first, it allows for the best wavelength(s) to be selected for actual analysis. This is particularly important when no information is available on molar absorptivities at different wavelengths.
The second major advantage is related to the problem of peak purity. Often, the peak shape in itself does not reveal that it actually corresponds to two (or even more) components. In such a case, absorbance rationing at several wavelengths is particularly helpful in deciding whether the peak represents a single compound or, is in fact, a composite peak.
In absorbance rationing, the absorbance is measured at two or more wavelengths and ratios are calculated for two selected wavelengths. Simultaneous measurement at several wavelengths allows one to calculate the absorbance ratio. Evaluation can be carried out in two ways:
In the first case, the ratios at chosen wavelength are continuously monitored during
the analysis: if the compound under the peak is pure, the response will be a square wave
function (rectangle),. If the response is not rectangle, the peak is not pure.