ABSTRAK Isolasi senyawa terpenoid dari ekstrak etil asetat kulit akar biduri (Calotropis gigantea) telah dilakukan. Kulit akar C. gigantea dimaserasi menggunakan pelarut metanol, kemudian dipartisi menggunakan pelarut n-heksana, dan etil asetat, sehingga diperoleh ekstrak pekat sebanyak 66,18, 20,45, dan 73,60 gram. Ekstrak etil asetat diisolasi menggunakan metode Kromatografi Kolom Cair Vakum (KKCV) sehingga diperoleh 5 fraksi gabungan, yaitu Fraksi A (0,25 g), B (0,62 g), C (0,30 g), D (2,75 g) dan E (5,80 g). Fraksi C direkromatografi menggunakan kromatografi kolom gravitasi menghasilkan 40 fraksi. Hasil uji fitokimia subfraksi C2 menggandung senyawa golongan triterpenoid dan karakterisasi menggunakan spektrometri FT-IR menunjukkan adanya gugus C-H, C=O, C=C, dan C-O (ester). Berdasarkan data spektrometri massa, diduga subfraksi C2 mengandung senyawa tarakseril asetat. Kata Kunci : Calotropis gigantea, tarakseril asetat, triterpenoid. ABSTRACT Isolation of terpenoid compound from ethyl acetate of Calotropis gigantea root bark. The root bark of C. gigantea was macerated using methanol solvent, then partitioned with n-hexane, and ethyl acetate, so obtained concentrated extracts of 66.18, 20.45, and 73.60 g. The ethyl acetate extract was isolated by Vacuum Column Chromatography method (VCC) so that obtained 5 fractions of combined, Fraction A (0.25 g), B (0.62 g), C (0.30 g), D (2.75 g) and E (5.80 g). Fraction C was recromatographed by gravity column chromatography obtained 40 fractions. The result of phytochemical test of subfraction C2 contains triterpene compound then characterized by FT-IR spectrometry showed the presence of C-H, C=O, C=C, and C-O (ester) groups. According to the characterization with mass spectrometry data, C2 subfraction expected have contained taraxeryl acetate compound. Keyword : Calotropis gigantea, taraxeryl acetate, triterpene.