Perioperative factors including hypoxia, hypocapnia and certain anesthetics have been suggested to contribute to Alzheimer’s disease (AD) neuropathogenesis. Desflurane is one of the most commonly used inhalation anesthetics. However, the effects of desflurane on AD neuropathogenesis have not been previously determined. Here, we set out to assess effects of desflurane and hypoxia on caspase activation, amyloid precursor protein (APP) processing and amyloid-ß protein (Aß) generation in H4 human neuroglioma cells (H4 naïve cells) as well as those overexpressing APP (H4-APP cells). Neither 12% desflurane nor hypoxia (18% O2), alone, affected caspase-3 activation, APP processing and Aß generation. However, treatment with a combination of 12% desflurane and hypoxia (18% O2) (desflurane/hypoxia) for six hours induced caspase-3 activation, altered APP processing and increased Aß generation in H4-APP cells. Desflurane/hypoxia also increased levels of ß-site APP-cleaving enzyme (BACE) in H4-APP cells. In addition, desflurane/hypoxia-induced Aß generation could be reduced by a broad caspase inhibitor Z-VAD. Finally, the Aß aggregation inhibitor clioquinol and -secretase inhibitor L-685,458 attenuated caspase-3 activation induced by desflurane/hypoxia. In summary, desflurane can induce Aß production and caspase activation, but only in the presence of hypoxia. Pending in vivo confirmation, these data may have profound implications for anesthesia care in elderly patients, and especially those with AD.