Background: The hepatoprotective effect of a polyherbal formulation was evaluated in female rabbits. The herbal formulation was used alone and in combination with ascorbic acid (AsAc) in animals with paracetamol (ParCM)-induced hepatic damage. Methods: The study design included five groups, each comprised of five animals. Group A was the control group (untreated; given only routine diet) while group B was given a single oral dose of ParCM (2 g/kg) on day 9. Groups C, D and E were pretreated with polyherbal formulation (PoHF; 500 mg/kg), ascorbic acid (AsAc; 200 mg/kg) and PoHF (500 mg/kg) combined with ascorbic acid (AsAc; 200 mg/kg), respectively for 9 consecutive days. On the last day (day 9), after 30 minutes of routine treatments, a single dose of ParCM (2 g/ kg) was administered in groups C, D and E. Animals were sacrificed 24 hours after the last treatment. Blood and liver samples were collected from all animals. Serum was separated from the blood samples and subjected to biochemical tests for liver biomarker analysis. The biomarkers included alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP) and total bilirubin. Elevation of enzyme markers was considered an indicator of hepatocellular injury. Results: Serum levels of liver enzymes and total bilirubin were elevated significantly in group B when compared to group A. The level of serobiochemicals significantly dropped in group C but increased significantly in group D, as compared to group B.