Context: Liver cancer is widespread liver malignancy in the world, for an estimated one million deaths annually. Objective: In present work, lactobionic acid conjugated PLGA nanoparticles (LDNPs) bearing 5-Fluorouracil (5-FU) were developed for targeted delivery to hepatocellular carcinoma. Materials and methods: Lactobionic acid conjugated PLGA was used to prepare LDNPs using modified emulsion diffusion method. Results: They were characterised for particle morphology, particle size (below 150 nm), zeta potential and polydispersity index (PDI similar to 0.35), entrapment efficiency (similar to 60.23%), and cumulative percent drug release. Discussion: LDNPs in ex-vivo cell line studies on human cancer cell line HepG2 exhibited significantly higher cytotoxicity compared to 5-FU and DNPs (unconjugated PLGA NPs) with growth inhibition 50% (GI(50)) of 66.7 mu g/mL, 50.2 mu g/mL and 35.5 mu g/mL, respectively. In vivo studies exhibited higher drug concentration about 37.52 +/- 0.68% in liver as compared to other organs and plasma. Conclusion: Thus, LDNPs showed high drug loading, specificity, biocompatibility and efficacy in treatment of liver cancer.