A stereospecific method for the analysis of propranolol and 4-hydroxypropranolol in human plasma employing fluorescence detection has been developed using the homochiral derivatizing agent 2,3,4,6-tetra-O-acetyl-beta-D-glucopyranosyl isothiocyanate (TAGIT). The use of fluorescence detection provided enhanced sensitivity and cleaner chromatograms for the analysis of plasma samples, when compared to UV detection. Furthermore, parameters such as TAGIT concentration, vortex time and reaction time were examined to optimize conditions for maximum derivatization recoveries. The analyses of S(-)- and R(+)-propranolol from plasma were linear over the concentration range of 2.0-200 ng ml-1, while S(-)- and R(+)-4-hydroxypropranolol were linear from 5.0 to 200 ng ml-1.