Determination of pteridines and flavins in seawater by reverse-phase,high-performance liquid chromatography with fluorometric detection

Methods are described for routine analyses of individual pteridines and flavins from marine waters at picomolar concentrations. Samples were concentrated by adsorption on bonded-phase, C₁₈ silica and Florisil Sep-Pak extraction columns connected in series. Sample extracts derived from Sep-Pak eluates were analysed by reverse-phase liquid chromatography with fluorometric detection. These methods proved sensitive for the quantitative identification of pterin, xanthopterin, isoxanthopterin, 6-biopterin, pterin-6-carboxylic acid, 6-hydroxymethylpterin, riboflavin, riboflavin-5′-phosphate, lumiflavin and lumichrome in reef waters at Davies Reef, Great Barrier Reef, Australia.Preliminary experiments were conducted to test the application of these methods to the measurement of pterin and flavin excretion from several reef-dwelling, benthic organisms. Results from these experiments suggest that the benthic community contributes these compounds to the coral reef environment.