Computer-aided strategies for archive retrieval and sensitivity enhancement in the identification of drugs by photodiode array detection in high-performance liquid chromatography

An archive retrieval algorithm for high-performance liquid chromatography with UV detection (HPLC-UV) has been developed for the first time for the rapid identification of spectra acquired by rapid-scanning photodiode array detection in HPLC. The algorithm is based on a database of spectra (normalised with respect to area), inverse files of key spectral features, a selective search window, with parabolic weighting factors and least-squares comparison of test and retrieved spectra. The performance of the library search system is demonstrated with respect to a small library of solutes, including: cortisone acetate, ethynyl estradiol, ethisterone, progesterone; caffeine, theobromine, theophylline and 8-chlorotheophylline; morphine and diamorphine; and cycloserine. Since photodiode array detection operates in the domains of both wavelength and time, to generate a matrix of (A,?,t) data, the optimum conditions for sensitivity enhancement by ensemble averaging in these domains have been examined. At a given observation wavelength, increase in the detector bandwidth (or 'diode bunching') yields a value of ?? which gives optimum sensitivity; this value is systematically related to the spectral bandwidth of the analyte. Sensitivity can also be optimised by varying the integration period in the time domain. Sensitivity can be further increased by combination of these instrumental optima, for which there is evidence of dependence on the particular instrument design. Response was found to be linear for detector bandwidths up to twice the optimum value. The comparative sensitivity of some commercially-available photodiode array detectors has been assessed relative to that of conventional detectors under strictly controlled conditions.