Objective: To study the chemical constituents of Atractylodis Macrocephalae Rhizoma. Method: The compounds were isolated and purified from ethyl acetate extract of Atractylodis Macrocephalae Rhizoma by using a combination of chromatographic techniques. Their structures were elucidated on the basis of thierphysico-chemical properties and spectroscopic data analyses. MTT method was used to assay the cytotoxicity agianst HeLa cells. Result: Sixteen compounds were isolated and purified from ethyl acetate extract of Atractylodis Macrocephalae Rhizoma and determined as 1-acetoxytetradeca-6E

12E-dien-8

10-diyn-3-ol (1)

3-acetoxytetradeca-6E

12E-dien-8

10-diyn-1-ol (2)

tetradeca-6E

12E-dien-8

10-diyn-1

3-diol diacetate (3)

14-acetoxy-12-senecioyloxytetradeca-2E

8Z

10E-trien-4

6-diyn-1-ol (4)

14-acetoxy-12-senecioyloxytetradeca-2E

8E

10E-trien-4

6-diyn-1-ol (5)

atractylenolide Ⅰ (6)

atractylenolide Ⅱ (7)

atractylenolide Ⅲ (8)

biatractylenolide (9)

juniper camphor (10)

taraxerylacetate (11)

3β-acetoxy-12-oleanen-11-one (12)

7-hydroxy coumarin (13)

octacosanic acid (14)

heptadecanoic acid (15)

7-α-hydroxyl-β-sitosterol (16). Conclusion: Among them

compounds 1

2

16 were isolated from the species firstly and compounds 12-15 were isolated from the genus of Atractylodes for the first time. Compound 6 showed good inhibitory activity against HeLa cells with IC50 values of 15.6μmol ·L-1.