Objective To determine the content of Atractylone, Atractylenolide Ⅰ, Atractylenolide Ⅱand Atractylenolide Ⅲin Atractylodis Macrocephalae Rhizoma stir-fried with wheat bran from different habitats by high performance liquid chromatogram method, and to evaluate the quality of Atractylodis Macrocephalae Rhizoma stir-fried with wheat bran from different habitats. Methods The separation was performed on Dikma Diamonsil C8 (150 mm×4.6 mm, 5 μm)column with mobile phase consisted of water-acetonitrile (gradient elution) at the flow rate of 1.0 mL/min. The detection wavelength was set at 220 and 275 nm, the column temperature was 30℃and injection volume was 20 μL. Results Under the above chromatographic conditions, Atractylone, Atractylenolide Ⅰ, Atractylenolide Ⅱand Atractylenolide Ⅲ showed good linear relationship in the ranges of 47.600-142.800, 1.304-3.912, 1.126-3.378, 1.740-5.220 mg/L.The average recovery was 99.98%, 99.63%, 99.15%, 99.79%, respectively, and the relative standard deviation (RSD)was 0.03%, 1.70%, 1.57%, 1.10%, respectively. Conclusion This method is well-separated, specific, reproducible,simple and feasible. It can be used for the quality control method of Atractylodis Macrocephalae Rhizoma stir-fried with wheat bran. The result of determination shows that the content of the four components in Atractylodis Macrocephalae Rhizoma stir-fried with wheat bran from Zhejiang Pan′an is relatively higher.