Erythrocyte concentrates for transfusion purposes represent a life-saving therapeutics of primary relevance in the clinical setting. However, efforts have been continuously proposed to improve safety and efficacy of long-term stored red blood cells. By means of liquid chromatography coupled with Q-TOF mass spectrometry, we were able to perform an untargeted metabolomics analysis in order to highlight metabolic species (i.e. low molecular biochemicals including sugars, lipids, nucleotides, aminoacids, etc.), both in red blood cells and supernatants, which showed fluctuations against day 0 controls over storage duration on a weekly basis. We could confirm and expand existing literature about the rapid fall of glycolytic rate and accumulation of glycolysis end products. A shift was observed towards the oxidative phase of pentose phosphate pathway, in response to an exacerbation of oxidative stress (altered glutathione homeostasis and accumulation of peroxidation/inflammatory products in the supernatant). The present study provides the first evidence that over storage duration metabolic fluxes in red blood cells proceed from pentose phosphate pathway towards purine salvage pathway, instead of massively re-entering glycolysis via the nonoxidative phase. This article is part of a Special Issue entitled: Integrated omics.
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