Punarnavashtak (PN) kwath is a classical Ayurvedic formulation mentioned in Ayurvedic literature "Bhaishyajyaratnavali" for hepatic disorders and asthma. Standardization and clinical trial to support its efficacy are lacking. So, in the present study, standardization of PN kwath was done by using biomarkers, gallic acid and berberine, and its hepatoprotective activity was evaluated by clinical study to rationalise the traditional use of this formulation. PN kwath was standardized by HPTLC (High performance thin layer chromatography) using gallic acid and berberine as biomarkers and was subjected to clinical study. For clinical study patients attending outpatient clinics, with an evidence of liver disease were included in the study. During the study period, patients who fulfilled inclusion criteria were randomly assigned. The recommended dose was 20 ml kwath daily for 8 weeks. All the patients underwent clinical examination and laboratory investigations for liver functions tests before the commencement of therapy. Thereafter, clinical assessments were done after 8 weeks of treatment. The results showed significant changes in liver functions tests [serum glutamate oxaloacetate transaminase (SGOT), serum glutamate pyruvate transaminase (SGPT), alkaline phosphatase (ALP), total bilirubin]. There was no report of adverse effects attributable to this formulation. Our results suggest that PN kwath showed significant hepatoprotective activity. Berberine and gallic acid were found to be 0.08 and 4.9%, respectively. Our results suggest that PN kwath showed significant hepatoprotective activity due to presence of various phytoconstituents and support its traditional uses in liver disorder.