A definitive method to determine adenine compounds simultaneously was established by introducing a new fluorescent reagent into high-performance liquid chromatography. Bromoacetaldehyde was the best reagent among the haloacetaldehydes examined. A quantitative reaction was obtained even for unstable ADP and ATP. A high resolution of adenine nucleotides was obtained using a column of Hitachi gel No. 3012-N. The method was applied to the measurement of cyclic AMP in urine, and ADP and ATP in brain and blood. Further, the sensitivity of the method was increased by a new fluorescence spectrophotometer constructed for micro-HPLC. Femtomole amounts of the adenine nucleotides were clearly separated.

Acetaldehyde↗

PubMed1980

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.

Adenine↗