Sensitive fluorimetry of adenine, its nucleosides and nucleotides

We improved our fluorimetric analysis of adenine compounds by high performance liquid chromatography(HPLC). Bromoacetoaldehyde was better than Chloro- or iodoacetoaldehyde as the reagent of fluorescent derivatization. A moderate reaction for unstable ADP and ATP was found and their fluorescent derivatives were separated by HPLC using Hitachi gel No. 3012-N. The method was applied to determine cAMP in human urines and a catecholamine receptor system. Further, a sensitive fluorescence spectrophotometer was developed. In this micro-HPLC, several ten femtomoles of the adenine compounds were separated.