The aim of the study was to evaluate the impact of environmental contaminants on oxidative stress biomarkers in hepatocytes of Indian estuarine water grey mullet, Mugil cephalus collected from unpolluted Kovalam and polluted Ennore estuaries. Initially, a comparison was made between the general water chemistry and environmental pollutants like heavy metals to identify the ecotype of the estuaries. Biomarker responses and bioaccumulation of metals were determined along with histological studies of fish hepatocytes to assess contamination impact. Water chemistry data with higher temperature, salinity, pH, biochemical oxygen demand, chemical oxygen demand and low dissolved oxygen and environmental pollutants with higher metal concentration (p<0.05) were observed at Ennore than Kovalam indicating its polluted environment. A significant increase (p<0.05) in lipid and protein oxidation markers, decrease (p<0.05) in thiol status and antioxidant enzyme activities were observed in the Ennore fish hepatocytes compared to Kovalam counterpart. Bioaccumulation of heavy metals was significantly higher (p<0.05) in fish from Ennore. Hepatocytes from Ennore also featured extensive lipid-type vacuolation, increased size and membrane disruption. All the findings highlighted the value of oxidative stress biomarkers and membrane disruption as the sensitive parameters of environmental pollutant contamination and their importance in biomonitoring of aquatic ecosystems. This is also the first such attempt reported at the cellular level from South India stressing the importance of biomarkers in biomonitoring programmes using fish hepatocytes as the model system.