Abstract Modern science and technology world provides many scientific advances for understanding of molecular basis of diseases. However, still we have very difficult to understanding the diseases pathogenesis and in the development of successful approaching for early diagnosis and therapeutic treatments. The proteomic approaches has provided many chances to identifying molecular marker proteins and therapeutic targets. i.e., The present study was carried out to identify the protein expression associated with carbon tetrachloride produced free radical mediated oxidative stress in Swiss albino mice by using advanced technology 2D-polyacrylamide gel electrophoresis and matrix-assisted laser desorption /ionization-time of flight mass spectrometry. The mice were separated into four different groups [Each = 6 mice]. Group I served as normal. Group II served as control mice administered with lycorine alone [5mg/kg]. For inducing hepatotoxicity animals of groups III–IV were administered orally [1mL/kg body weight] with carbon tetrachloride twice a week for 8 weeks. Group III served as CCl4 induced group of mice. Group IV administered with lycorine [5mg/kg i.p] daily for 8 weeks. The proteins changes were observed by 2D gel electrophoresis which is given new information about metabolic and signaling pathways during CCl4 induction. The three differential protein expressions were excised from the liver of CCl4 induced and lycorine treated group of mice. The expressions of these proteins were further confirmed by MALDI-TOF-MS i.e. ATP synthase, regucalcin and HSP60. These identified proteins involved in regulation of ATP production, calcium regulation and maintaining the integrity of cellular proteins respectively. The results provide better elucidation of molecular mechanism of protein in liver function associated with free radicals related diseases.