The aim of the present investigation is to explore the innovative platform for the synthesis of plant-based nanoparticles, which contain biocompatible and biodegradable carrier of chitosan loaded with phloretin hydrophobic phytochemical applied as a stable anticancer agent. Treatment of cancer uses chemotherapeutic drugs as the cells are resistant to other drugs. However, the usage of therapeutic drug is limited by its poor solubility and low bioavailability. To overcome this problem, we fabricated the phloretin loaded chitosan nanoparticles (PhCsNPs) and physicochemical properties of PhCsNPs were characterized by FTIR, XRD, DLS, SEM and TEM. The findings indicated that the synthesized PhCsNPs were spherical and homogeneous in shape with the size distribution of 80-100 nm and exhibited stability in ultimate drug releasing profile. Further, we substantiated the anticancer efficiency of PhCsNPs through bio-assessment, such as cytotoxicity measurement, intracellular ROS, mitochondrial dysfunction, lipid peroxidation measurement, antioxidants status, apoptotic associated gene expression profile and cell cycle analysis in human oral cancer cell lines. The findings suggested that PhCsNPs augmented the mitochondrial-mediated apoptotic mechanism through the stimulation of oxidative stress, depletion of cellular antioxidants and cell cycle arrest. Our data suggested that PhCsNPs could be used as an efficient therapeutic agent for the treatment of oral cancer. © 2019