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Full metabolomics analyses revealed alterations in purine and arginine pathways at baseline, along with activation of pentose phosphate pathway and glycolysis upstream to pyruvate kinase in βThal+ RBCs. Upon storage, substantial changes were observed in arginine, purine and vitamin B6 metabolism, as well as in the hexosamine pathway. A high degree of glutamate generation in βThal+ RBCs was accompanied by low levels of purine oxidation products (IMP, hypoxanthine, allantoin). The βThal mutations impact the metabolism and the susceptibili