Hepatitis C virus (HCV) infection of the liver is a global health problem and a major risk factor for the development of hepatocellular carcinoma (HCC). Sensitive methods are needed for the improved and earlier detection of HCC, which would provide better therapy options. Metabolic profiling of the high-risk population (HCV patients) and those with HCC provides insights into the process of liver carcinogenesis and possible biomarkers for earlier cancer detection. Seventy-three blood metabolites were quantitatively profiled in HCC (n = 30) and cirrhotic HCV (n = 22) patients using a targeted approach based on LC-MS/MS. Sixteen of 73 targeted metabolites differed significantly (p < 0.05) and their levels varied up to a factor of 3.3 between HCC and HCV. Four of these 16 metabolites (methionine, 5-hydroxymethyl-2′-deoxyuridine, N2,N2-dimethylguanosine, and uric acid) that showed the lowest p values were used to develop and internally validate a classification model using partial least squares discriminant analysis. The model exhibited high classification accuracy for distinguishing the two groups with sensitivity, specificity, and area under the receiver operating characteristic curve of 97%, 95%, and 0.98, respectively. A number of perturbed metabolic pathways, including amino acid, purine, and nucleotide metabolism, were identified based on the 16 biomarker candidates. These results provide a promising methodology to distinguish cirrhotic HCV patients, who are at high risk to develop HCC, from those who have already progressed to HCC. The results also provide insights into the altered metabolism between HCC and HCV.