Plasmodium falciparum is the parasite responsible for causing the most severe form of human malaria. The disease manifests when P. falciparum invades and replicates in red blood cells (RBCs). During development in RBCs, the parasite secretes small molecules, proteins and vesicles into the extracellular environment. These extracellular factors have been found to activate host immune and endothelial cells in vitro. However, biochemical changes in bystander, uninfected RBCs (essential for the continuation of the parasite lifecycle) have not been described. We performed untargeted metabolomics to investigate the metabolism of uninfected RBCs exposed to media from a P. falciparum culture (“conditioned unRBCs”) compared to control unRBCs. We observed significant perturbations to glycolysis in conditioned unRBCs, with few significant changes in other metabolic pathways. To evaluate active glycolytic flux in conditioned unRBCs, stable isotope labelled glucose (U-13C-glucose) was spiked into the cultures 3 h prior to metabolite extraction. Our data showed that 13C-metabolite isotopologues were depleted upstream of pyruvate kinase (a rate-limiting enzyme of glycolysis) and accumulated downstream of this enzyme, reflecting an increase in enzyme activity. Furthermore, the levels of 13C-lactate secreted by conditioned unRBCs were found to be higher than control cells, indicating that extracellular factors from P. falciparum increase the glycolytic flux of unRBCs. To identify the parasite factors driving this effect, media from a P. falciparum culture was filtered (to remove protein and extracellular vesicles) and incubated with unRBCs. Our results showed that the remaining parasite small molecules, alone, were capable of causing glycolytic perturbations to unRBCs. Metabolomic profiling of media from P. falciparum cultures has provided a candidate list of small molecules that may be responsible for this effect. Altogether, these data demonstrate that P. falciparum extracellular factors impact unRBC metabolism, which may contribute to the metabolic symptoms of malaria such as hypoglycaemia, lactic acidosis and anaemia.