Hepatocellular carcinoma (HCC) is a subtype of liver cancer with high worldwide prevalence and mortality. Its high recurrence rate and heterogeneity have challenged effectiveness of existing therapies. Numerous researchers explore the multifaceted benefits of Vitamin C (VC) in cancer treatments. In this study, we utilized the NMR-based metabolomics approach to systematically surrogate the molecular basis underlying the anticancer property of VC in HCC cell. Moreover, combination of VC with chemotherapeutic agent was tried to investigate synergetic effects in modulating metabolic profiles of cancer cells. Our results help to reveal the molecular basis of HCC treatment and may facilitate future clinical therapy designs.