HPLC-DAD coupled to an electrospray tandem mass spectrometer (ESI-MS) has been developed to analysis the chemical profile of Danning tablets. Separation was performed on a ZORBAX Extend C18 analytical column by gradient elution with acetonitrile and formate buffer (containing 12.6 mmol L−1 formic acid, adjusted to pH 5.0 with ammonia) at a flow rate of 0.8 mL−1 min. Twenty-three peaks were identifies as the major compounds in this preparation. Similarity measurements of the HPLC-UV chromatograms were investigated by a Similarity Evaluation System for Chromatographic Fingerprinting of Traditional Chinese Medicine (TCM) in the range of 0.84–0.99, and principal components analysis (PCA) on the quantitative analysis data was further performed to evaluate the variation among batches. The proposed method gives chemical profiles of multi-components in one run, and therefore can be readily utilized as a comprehensive quality control approach for TCM.