Sibrowski W, Lüdemann K, Dominka T, von Eisenhart-Rothe B, Kühnl P
Abt. f. Transfusionsmedizin u. Transplantationsimmunologie, Universitäts-Krankenhaus Eppendorf, Hamburg.
Beitr Infusionsther. 1990;26:86-8.
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.
我们采用反相高效液相色谱法（HPLC）开发了一种快速测定人红细胞（RBC）中腺嘌呤核苷酸ATP、ADP和AMP的方法。该方法灵敏度高（10⁻⁹ - 10⁻¹⁰ M）、特异性强（峰纯度大于95%），结果重现性良好。这三种物质可一步测定，这对于准确测定腺苷酸能量电荷（AEC）至关重要。我们将该技术应用于新鲜和储存的红细胞。在PAGGS - 山梨醇中储存六周后，我们发现ATP含量仅略有下降，但ADP和AMP含量显著增加。相应地，AEC降低，而总核苷酸池（TNP）没有明显变化。由于AEC和TNP反映细胞能量代谢状态，它们与储存红细胞的活力有关。因此，我们认为所描述的方法有助于对储存在新的或改良培养基中的红细胞进行质量控制。
