Measurement of red blood cell adenosine nucleotides by high-performance liquid chromatography

A new method for measuring ATP in stored red blood cells using high-performance liquid chromatography was compared with an established enzymatic method. The new method is based upon isocratic reverse phase chromatography using a polyvinyl alcohol gel stationary phase. The chromatograms produce quantitative results for ADP, AMP, and other nucleotides, and can be used to determine adenylate energy charge. The correlation coefficient between the two methods was 0.91, and mean ATP was 3.0 mumol/g Hb for both methods. Tests of hypothesis for mean and variance were not significant. The method is recommended as a means to study the relationships between poststorage red blood cell ATP, adenylate energy charge, total adenylates, and posttransfusion erythrocyte survival.