Quality control and screening of active substances in traditional Chinese medicines have been performed using ifngerprint analysis. The spectrum-effect relationship between chromatography ifngerprints and efifcacy of herbal drugs is considered as a potentially useful method for determining active ingredients in complex mixtures. The study was designed to develop a method for determining the bioactive components of a compound Chinese medicine called Tiefeng based on spectrum-effect relationships between high-performance liquid chromatography (HPLC) ifngerprints and anticoccidial activities. Four peaks of the established HPLC ifngerprint indicate the main bioactive components of this medicine. In addition, pharmacodynamic atlas was deifned and used to assess the anticoccidial activity of Tiefeng from different sources for the ifrst time. We found that the level of anticoccidial activity of Tiefeng was consistent with the degree of similarity between the pharmacodynamic atlas and chromatogram of any sample. Furthermore, effect of this medicine was related with the main active constituents, along with the origin and the harvesting time.