The pyrrolophenanthridine alkaloid lycorine has frequently been used as a specific inhibitor to help elucidate the function of ascorbic acid (vitamin C) in a wide range of biological processes. It was recently reported that this function can be exercised by inhibiting the activity ofl-galactono-1,4-lactone dehydrogenase, the terminal enzyme of ascorbic acid biosynthesis, although working with the purified enzyme, we have been unable to repeat this result. Here, we present a procedure for the purification and analysis of lycorine by high-performance liquid chromatography from twoCrinumspecies and describe for the first time a method that allows the simultaneous analysis of ascorbic acid and lycorine in tissue extracts ofCrinum asiaticumby micellar electrokinetic chromatography.