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Objective: To establish a convenient and economic method to determine hepatoma-specific α-fetoprotein (HS-AFP) for diagnosis of hepatocellular carcinoma (HCC). Methods: HS-AFP from serum of HCC patients was separated by a mini-column Lens culinaris agglutinin (LCA)-affinity chromatography. The levels of serum total AFP and separated HS-AFP were detected by radioimmunoassay (RIA). Results: Circulating AFP was separated into three peaks (AFP-1, AFP-2, and AFP-3) by LCA-affinity chromatography. During the elution course, the AFP-1 and AFP-2 could be eluted with TE buffer. HS-AFP (AFP-3) from sera of HCC patients was eluted dearly on the LCA-sepharose gel mini-column with a solution containing a-methyl-D-mannoside. It was a part of total AFP and only found in sera of HCC patients. A ratio of more than 15% for HS-AFP to total AFP in serum was considered as a specific marker for HCC diagnosis with higher sensitivity (92.7%) and specificity(88.2%). Conclusion: The new assay for circulating HS-AFP analysis is more sensitive, repeatable, and convenient. Its clinical application would be useful to early diagnosis of HCC.