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This Small Business Innovation Research (SBIR) Phase II project will lead to a breakthrough in the use of laser-induced fluorescence (LIF) for chromatographic detection. Commercial standalone LIF detectors are based on CW lasers and collect data at a fixed wavelength. Consequently, they add minimal capability for resolving complex mixtures beyond that inherent in the chromatographic separation itself. On-the-fly fluorescence lifetime measurements at a single emission wavelength have been proposed as a better way to resolve the signals of co-eluting species. Our approach is far more powerful because it provides lifetimes on-the-fly and at several wavelengths simultaneously. A new prism flow cell fiber optically coupled to the emission spectrograph was introduced in Phase I. In addition, two different algorithms strategies for analyzing the multi-dimensional fluorescence data were developed and demonstrated. In Phase II a diode-pumped laser will replace the flashlamp pumped excitation laser, thereby providing 100 times higher pulse repetition frequency, 10 times shorter pulse duration, and 10 times better shot-to-shot stability. New digitizer technology will be incorporated to accommodate the laser's high repetition frequency. Important Phase II activities include fluorescence methods development to extend the range of applications to drugs and drug metabolites and elaboration of the chemometric algorithms. The instrumental approach to be realized through the Phase II research will have a profound impact on QA/QC assessments of drug purity, bioequivalence and pharmacokinetic studies, and research investigations in humans and animals. Sales of several hundred units per year to pharmaceutical manufacturers, contract research organizations, and universities are anticipated. The technology will later be adapted for faster and more accurate DNA sequencing