Sensitive fluorimetry of adenine-containing compounds with high-performance liquid chromatography.
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.
D000079:Acetaldehyde / Q000031:analogs & derivatives; D000225:Adenine / Q000032:analysis*; D000227:Adenine Nucleotides / Q000032:analysis / Q000097:blood; D000244:Adenosine Diphosphate / Q000032:analysis; D000249:Adenosine Monophosphate / Q000032:analysis; D000255:Adenosine Triphosphate / Q000032:analysis; D000818:Animals; D001923:Brain Chemistry; D055598:Chemical Phenomena; D002621:Chemistry; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D000242:Cyclic AMP / Q000652:urine; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D051381:Rats; D011919:Rats, Inbred Strains; D013050:Spectrometry, Fluorescence / Q000379:methods
D000227:Adenine Nucleotides; C038235:bromoacetaldehyde; D000249:Adenosine Monophosphate; D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate; D000242:Cyclic AMP; D000079:Acetaldehyde; D000225:Adenine