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.
Yoshioka
M
M
Faculty of Pharmaceutical Sciences, Setsunan University, Osaka, Japan.
Yamada
K
K
Abu-Zeid
M M
MM
Fujimori
H
H
Fuke
A
A
Hirai
K
K
Goto
A
A
Ishii
M
M
Sugimoto
T
T
Parvez
H
H
D000225:Adenine / Q000097:blood; D000227:Adenine Nucleotides / Q000032:analysis*; D000241:Adenosine / Q000032:analysis*; D000244:Adenosine Diphosphate / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000378:metabolism; D000262:Adenylyl Cyclases / Q000032:analysis; D000818:Animals; D002478:Cells, Cultured; D002851:Chromatography, High Pressure Liquid; D004910:Erythrocyte Membrane / Q000032:analysis; D006801:Humans; D051379:Mice; D009447:Neuroblastoma / Q000378:metabolism; D010042:Ouabain / Q000494:pharmacology; D051381:Rats; D000254:Sodium-Potassium-Exchanging ATPase / Q000037:antagonists & inhibitors; D013050:Spectrometry, Fluorescence
D000227:Adenine Nucleotides; D010042:Ouabain; D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate; D000262:Adenylyl Cyclases; D000254:Sodium-Potassium-Exchanging ATPase; D000225:Adenine; D000241:Adenosine