Glycofurol (GF) has been used clinically as a solvent for parenteral drug delivery systems. However, the application and toxicity of GF in the brain have not been reported. This study was carried out to assess the systemic and neurologic reactions of GF in rats upon intracranial injection. Hematological and neuropathological assessments of rats were performed during the acute, subacute and chronic period after the injection. Injection of the GF solution (GF 25 μL + PBS 25 μL) into the brain cortex showed that it did not cause any deaths or clinical neurobehavioral abnormalities. At the same volume as phosphate-buffered saline (PBS) injection, it had mild effects on all hematological data and histopathology of brain tissues. Nevertheless, histomorphologic assessments of the brain tissues treated with PBS 70 μL revealed different tissue responses compared with those of 70 μL GF solution (30 μL + PBS 40 μL) where tissues around the administration site showed elevated polymorphonuclear leukocytes, macrophages and gliosis. These results demonstrated that the GF solution (GF 25 μL + PBS 25 μL) administration was well tolerated and caused minor inflammatory responses of cerebral cortex. This suggests possibilities of GF for drug delivery systems in the brain parenchymal tissues.