LASER-INDUCED FLUORESCENCE DETECTION OF NATIVE-FLUORESCENT ANALYTES IN COLUMN LIQUID-CHROMATOGRAPHY, A CRITICAL-EVALUATION

This paper critically evaluates aspects that need to be considered whe n applying lasers instead of conventional lamps in fluorescence detect ion combined with column liquid chromatography (LC). To exclude the of ten underestimated role of chemical derivatization reactions as far as detection limits are concerned, emphasis is on analytes showing nativ e fluorescence. First of all basic features of both continuous wave an d pulsed lasers are considered, including a quantitative treatment of saturation and photodecomposition effects. Subsequently a detailed tre atment on signal-to-noise in laser induced fluorescence (LIF) is prese nted, the various definitions of detection limits operative in the lit erature are compared and flow cell constructions are considered. The p aper ends with a comparison between conventional fluorescence and LIF detection in LC, an overview of published data on detection limits and a discussion on future trends.