No single biologic marker is used in the routine diagnosis of hepatocellular carcinoma (HCC). We screened for potential biomarkers in 142 samples, including samples from 60 patients with HCC, 36 patients with hepatic disease other than HCC, and 46 age- and sex-matched healthy volunteers. Pretreated samples were analyzed using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) combined with weak cationic exchange magnetic beads. The diagnostic pattern with a panel of 4 potential protein biomarkers of a mass-to-charge (m/z) ratio of 4,471, 8,936, 11,670, and 13,752 could accurately recognize 56 of 60 patients with HCC, 34 of 36 patients with hepatic disease, and 44 of 46 healthy people. The preliminary data suggest a potential application of MALDI-TOF MS combined with magnetic beads as an effective technology to profile serum proteome, and, with pattern analysis, a diagnostic model comprising 4 potential biomarkers was indicated to differentiate people with hepatic disease and healthy control subjects rapidly and precisely.