Objective:To establish a surface enhanced Raman spectroscopy ( SERS) technique for the determination of atropine sulfate. Methods:The Raman peaks of atropine sulfate molecules were classified by theoretical calculations and experimental tests, and the 1002 cm-1 Raman peaks were selected as the characteristic peaks for the quantitative analysis. Results:The detection limit of atropine sulfate in aqueous solution was below 0. 5 μg·ml-1 . The relationship between the intensity of characteristic peaks at 1002 cm-1 and the concentration of aqueous solution was linear within the range of 1-8 μg ml-1 with the linear correlation coefficient r of 0. 9839. The recovery rates of 2, 5 and 7μg ml-1 were measured, which were 97. 1%-109. 8%. The average recovery was 103. 3%, and the RSD was 4. 5% (n=9). At the same time, the stability of the method among the batches was tested, and the relative standard deviation was 5. 7 %. In addition, the atractylodes rhizome samples containing atropine sulfate were detected,and the characteristic peaks still could be detected at 1002 cm-1 . Conclusion:The method is rapid, accurate and nondestructive with easy operation, which can be used for the detection of atropine sulfate.