This study aimed to evaluate the effect of Hibiscus sabdariffa L. dried calyx ethanolic (HSE) extract on the redox imbalance and glucose plasma of vitamin B12 restriction Sprague-Dawley rats. The rat was fed a Vitamin B12 Restriction Diet for 16 weeks and treated with HSE as the treatment group. One group was fed a restriction diet not treated with HSE as a positive control group, and another was fed a control chow diet not treated with HSE as a negative control group. Vitamin B12, MDA, SOD activity enzyme, and glucose levels were evaluated in this study. The extract had a significant antioxidant capacity in terms of reducing the production of free radical scavenging activity. The HSE can repair the skewed redox imbalance and oxidative damage generated by a vitamin B12-deficient diet in the rat liver. The result of fasting glucose plasma levels, in 16 weeks showed both significantly different in all groups. In Sprague-Dawley rats with vitamin B12 restriction treated with HSE, a decrease in the glucose plasma in week sixteen was observed, together with a reduction of hepatic redox imbalance. Treatment with HSE protected hepatocytes from oxidative damage caused by vitamin B12 deficiency.