A microbore liquid chromatography (microbore LC) has been developed for the simultaneous determination of atractylenolide III, atractylenolide I and atractylon in Artactylodis macrocephala. In the method, a Microsil C18 column (150 mm x 1.0 mm) was used with the mobile phase containing methanol-acetonitrile-water for gradient elution, the flow rate was 50 microL/min, and the detection wavelength was set at 220 nm. Under the optimized separation conditions, every component was separated thoroughly. The relationships between the concentrations and the peak areas of the three components were all linear. The recoveries were 96.86% for atractylenolide III, 97.13% for atractylenolide I and 98.06% for atractylon, the relative standard deviations (RSD) were 1.63%, 1.31% and 0.39%, respectively. The method is simple, convenient, and can be used for the quality control of Artactylodis macrocephala.