Vitamin C is crucial for the brain. We aimed to investigate the effects of vitamin C administration following 24 hours of acute food deprivation and 24 hours of acute food intake on changes in vitamin C levels in different brain areas of guinea pigs. Vitamin C was administered as a single intraperitoneal dose (500 mg kg-1 body weight) both before acute food deprivation and before acute food intake. At the end of our study, we measured the vitamin C levels in cerebral cortex lobes, brain stem structures, hypophysis, hypothalamus, cerebellum, hippocampus, and amygdala. Vitamin C levels in the frontal and parietal lobes were found to be significantly higher in animals pretreated with vitamin C prior to 24 hours of food deprivation (p < 0.05). Temporal lobe vitamin C level was significantly lower in animals that were subjected to 24 hours of acute food intake following 24 hours of food deprivation (p < 0.05). Increased vitamin C levels were observed in the occipital lobe of all animals that received vitamin C administration (p < 0.05). Vitamin C levels in the brain stem structures such as mesencephalon and pons were significantly decreased in animals pretreated with vitamin C before normal feeding (p < 0.05). Vitamin C level in the hypothalamus was significantly increased after 24 hours of food deprivation (p < 0.05). In conclusion, different areas of the brain may differ in terms of vitamin C content during nutritional changes with or without vitamin C pretreatment, such as 24 hours of food deprivation or 24 hours of food intake following 24 hours of food deprivation. These differences may be attributed to several functions of vitamin C which may occur under these circumstances.