Analyses of adenosine and adenine nucleotides in biological materials by fluorescence reaction-high-performance liquid chromatography.

A previous method of determination of adenine compounds by high-performance liquid chromatography, using bromoacetaldehyde as a fluorescent reagent and a column of Hitachi gel No. 3012-N, was improved and extended to biological materials, especially to measure enzyme activities. A column packed with finer beads, Hitachi gel No. 3013-N, was found to be better than that of No. 3012-N, judging from the analysis time and resolution. ADP, from the hydrolysis of ATP by Na, K-ATPase, was determined quantitatively, and the enzyme activity was inhibited with ouabain. cAMP obtained from ATP by reaction with adenylate cyclase was also determined in the presence of various concentrations of L-epinephrine or sodium fluoride. The ATP levels in human blood were determined, and the cellular levels of ATP and ADP in neuroblastoma N1E 115 were examined as a function of cell growth.