Recent methods for determination of thiamine (thiochrome) and riboflavin by liquid chromatography (LC) are outlined and discussed, and a new method allowing the simultaneous determination of these 2 vitamins by using a single fluorescence detector is described. This system involves an ODS 5 micron ultrasphere column and a pH 7.5 mobile phase composed of 0.005M tetrabutyl ammonium phosphate in methanol-water (20 + 80).