[Rapid and sensitive determination of the adenylate energy charge in stored erythrocytes using high pressure liquid chromatography]. / Schnelle und sensitive Bestimmung des Adenylat-Energy-Charge in gelagerten Erythrozyten mittels Hochdruck-Flüssigkeits-Chromatographie (HPLC).

ABSTRACT

Using reversed-phase high-performance liquid chromatography (HPLC), we developed a fast method for the determination of the adenine-nucleotides ATP, ADP, and AMP in human red blood cells (RBC). The method is sensitive (10(-9)-10(-10) M), specific (greater than 95% peak-homogeneity), and the results are well reproducible. The three substances are assayed in a single step, which is essential for the exact determination of the adenylate-energy-charge (AEC). We applied the technique to fresh and stored RBC's. After six weeks' storage in PAGGS-sorbitol we found only a slight decrease in ATP content, but marked increases in ADP and AMP contents. Accordingly, there was a decrease in the AEC, whereas the total nucleotide pool (TNP) did not change significantly. As AEC and TNP reflect the state of the cellular energy metabolism, they are related to the viability of stored RBC. Thus, we consider the described method to be helpful for the quality control of RBC stored in new or modified media.