Microsomal cytochrome P450 is an important enzyme involved in drug metabolism and bioactivation in the liver. In guinea-pigs, the specific amount of cytochrome P450 depends on dietary vitamin C intake. Short-term vitamin C treatment can induce non-genomic effects on hepatocytes, such as activating mitochondrial respiration without changing cytochrome concentration. The aim of this investigation was to elucidate the long-term effects of a low or high vitamin C diet on the quantity and size of mitochondria in the hepatocytes of guinea-pigs. After 6 - 8 weeks of vitamin C feeding, the group receiving a low vitamin C content in their food (10 mg/100 g food) had significantly less mitochondria, but of a larger size, than the group receiving a high vitamin C content (680 mg/100 g food). This animal model investigation demonstrated that treatment with long-term vitamin C can induce morphological changes in the mitochondria of hepatocytes.