High Performance Liquid Chromatography/Video Fluorometry. Part I. Instrumentation.

A rapid scanning, two-dimensional fluorometer has been used to obtain Emission-Excitation Matrices EEMs of the effluent of an HPLC. These spectra, obtained on the fly, provide qualitative as well as quantitative information about unknown samples. An in-depth discussion of the instrumentation developed as well as data processing alternatives is presented. Current detectors for HPLC have certain limitations in the ability to completely characterize the column effluent during the separation of a complex sample. These limitations result, chiefly, from a lack of selectivity in the detectors used. One such example is the refractive index RI detector which monitors the change in a bulk property of the effluent, the refractive index. Even absorption and fluorescence detectors which use a single monitoring wavelength are typically more selective and sensitive than RI detectors. Still, the multicomponent capabilities of these devices are severely limited. More selective detection schemes have been developed. These employ atomic absorption and electrochemical techniques to selectively analyze components. Advanced, scanning absorption and fluorescence detectors have improved multicomponent capabilities.