Analysis of Purines and their Nucleosides by the Reverse Phase Partition Mode of High Pressure Liquid Chromatography
Although the analysis of nucleotides in blood is now being carried out routinely by high pressure liquid chromatography (HPLC) (1–4), and work is in progress to determine alterations casued by various disease states of free nucleotide concentrations in physiological fluids and cell extracts (5,6), less attention has been paid to free nucleoside levels in cells. Recent investigations, however, have focused attention on methylated nucleosides in cells as biological markers in cancer (7,8). Moreover, nucleoside concentrations are thought to be important in cardiac disease and birth defects. The intracellular level of adenosine, one of the physiological regulators of coronary blood flow, has been found to increase in cardiac hypertrophy and after brief periods of myocardial ischemia and hypoxia (9,10). In abnormalities in purine metabolic pathways caused by disease, the concentrations of purine bases and/or their nucleosides often are involved. For example in patients with adenosine deaminase deficiency, it has been observed that excess adenosine in cultured mammalian cells is toxic (11) and it has been postulated that accumulated adenosine is associated with severe immunological defects in children (12).
KeywordsXanthine Oxidase High Pressure Liquid Chromatography Purine Base Potassium Dihydrogen Phosphate Partition Mode
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