Objective To establish the chromatographic conditions of isochlorogenic acid A and isochlorogenic acid C in vernonia anthelmintica. Methods By changing the mobile phase,flow rate,column temperature and other chromatographic conditions,the best chromatographic conditions was we pursued to established. Results The linear relationship between the concentration of isochlorogenic acid A and the peak area was between 5. 825-69. 9 μg·mL-1, and the concentration of isochlorogenic acid C,was between 5.15-61.80 μg·mL-1and the peak area was good . The sample recovery rates of the two groups were 98.70%-101.92%(RSD＝1.04%,n＝9)、95.99%-102.52%(RSD＝1.90%,n＝9). Conclusion The method is simple,rapid, accurate and reliable for the determination of isochlorogenic acid A and isochlorogenic acid C in Vernonia anthelmintica and also for the quality control of the raw material.