The AcOEt eluate (45.8 g) was repeatedly chromatographed over silica gel with CHC13-MeOH and benzene-AcOEt to give betulonic acid as colorless powder (424 mg). Determination of antihepatotoxicity Assays using CCI4-and GaIN-induced cytotoxicity in primary cultured rat hepatocytes in vitro were carried out as described previously (11, 12).