Hesperetin (HSP) is a natural bioflavonoid found abundantly in citrus fruits with promising antioxidant, anti-inflammatory, anti-hypertensive andanti-cancer properties. The aim of the present study was to evaluate the in vivoanticancer activityof synthesized HSP loaded PLGA nanoparticles (PLGA-HSP NPs) in Ehrlich Ascites Carcinoma (EAC) cell bearing Swiss albino miceusing the standard drug 5-fluorouracil. PLGA-HSP NPs led to a considerable reduction in tumor volume, the number of viable tumor cells, and body weight in EAC tumor-bearing mice, as well as an increase in the mean survival time. PLGA-HSP NPs protect the host tissues from oxidative stress by recovering the antioxidant status in host mice. PLGA-HSP NPs also restored the haematological parameters. The aforementioned finding revealed that PLGA-HSP NPs have a favourable impact on EAC cell bearing Swiss albino mice. Therefore, the findings of the present study indicate that PLGA-HSP NPs may be considered as a competent anticancer agent in future.